Celebrating Prof. Edwards’ Receiving the Nobel Prize

16th World Congress on
In Vitro Fertilization
6th World Congress on In Vitro Maturation

September 10-13, 2011
Tokyo, Japan

Final Program and Abstracts

International Society for In Vitro Fertilization
with the cooperation of

The Japan Society of Fertilization and Implantation
Congress President: Osamu Kato (Director, Kato Ladies Clinic)
Congress Vice-President: Hisao Osada (Former Professor, Nihon University)TABLE OF CONTENTS■FINAL PROGRAM
3
Welcome Messages
7
Committees
9Congress Information
10
Date and Venue, Contacts, Registration, Message Board, Poster Area
11
Commercial Exhibition, Lunch and Coffee
12Instructions for Speakers and Chairpersons
14
Instructions for Poster Presenters
15
Floor Plan of the Congress Venue
16
Social Program and Travel Desks18
Map of the Congress Venue
19
Access to the Congress Venue
20
Airport Limousine Bus Time Table
21
Local Information
27
Agenda-at-a-Glance
31
Announcement of the 17th World Congress on In Vitro Fertilization, Tunis, Tunisia 201332
Scientific Program
33
1. Special Guest Lecture33
2. Opening Ceremony, Opening Lecture and Welcome Reception
33
3. Plenary Lectures
34
4. Pre-Congress Workshops
35
5. Concurrent Symposia
44
6. STGO Session
45
7. ISF Session
45
8. APART Session
46
9. Oral Communications
51
10. Poster Presentations
60
11. Luncheon Seminars■ABSTRACTS
62
Special Guest Lecture
65
Pre-Congress Workshops
75
Plenary Lectures
90
Concurrent Symposia
219
Society Sessions
238
Oral Communications
260
Poster Presentations
305
Author Index
306
Special Guest Lecture, Pre-Congress Workshops, Plenary Lectures, Concurrent Symposia
309
Oral Communications and Poster Presentations■CERTIFICATE
317
Certificate of Attendance (Copy)

2

WELCOME MESSAGESWELCOME MESSAGE FROM THE PRESIDENT OF ISIVF
Dear Colleagues,
The 16th World Congress on In Vitro Fertilization (IVF), which will be held in TokyoJapan in September 2011, is the main International Meeting of the year focusing on IVF
and Assisted Reproductive Technologies (ART). With the rapid advances that continue
to be achieved in these fields, this congress will constitute an important and uniquemultidisciplinary forum for the exchange of knowledge and the development of new ideas between the clinicians,
basic and clinical scientists, embryologists and allied health professionals, that it brings together.
The congress is being held in the exciting, dynamic, progressive yet traditional city of Tokyo. Come and enjoy
the congress and all of the historic, cultural, artistic and culinary delights that this most welcoming and uniquecity has to offer.
I would like to warmly thank the Japanese team and especially Dr Hisao Osada who has organized the 16th
World Congress on IVF despite great difficulties related to the recent tsunami and its dramatic consequences.
All the speakers have agreed to come and the participants will be numerous both because of the quality of thescientific program and also to show our solidarity.
I wish you a wonderful Congress and a memorable stay in Tokyo.

René FRYDMAN
President, International Society for In Vitro Fertilization

ABOUT ISIVF
ISIVF‟s primary mission is to promote research and clinical development of in vitro fertilization (IVF) of humanoocytes for the treatment of infertility and as an Assisted Reproductive Technology. Objectives include:
1. Promulgating ethical practice and standards of practice of IVF treatment;
2. Fostering collaboration between the various centers in the world that offer IVF treatment;
3. Organizing symposia for the purposes of presentation, discussion, exchange and publication of research, data, knowledge,
theories and standards pertaining to IVF, and specially, a forum under the name “World Congress on In Vitro Fertilization”.

3

WELCOME MESSAGESWELCOME MESSAGE FROM THE VICE-PRESIDENT OF ISIVF
Dear Colleagues,
It is with great pleasure that I welcome you to this 16th World Congress of In Vitro
Fertilization and to one of the world‟s most interesting and exciting cities: Tokyo.
This congress celebrates Professor Robert G Edwards and his great accomplishments
for which he has been awarded the Nobel Prize. This prestigious award honors him; it also honors the whole
field of reproductive medicine which has been transformed by him.
Holding the Congress such a short time after the devastating earthquake and tsunami, Japan experienced, is atestimony to the resilience of the Japanese people and the strong commitment and hard work of the localorganization headed by the President and Vice president of the Congress Dr. Osamu Kato and Prof. Hisao
Osada. I thank them personally and on behalf of the International Society for In Vitro Fertilization.
I trust that you will enjoy the rich and interesting academic and social program the congress offers. I also hopethat you will take the opportunity to enjoy Tokyo and other parts of Japan, and its unique and exquisite art andculture.

Victor GOMEL
Vice-President, International Society for In Vitro Fertilization

4

WELCOME MESSAGESWELCOME MESSAGE FROM THE PRESIDENT OF IVM
Dear Colleagues,
I am pleased to welcome you to the 6th World Congress on IVM and to Tokyo.
On behalf of the International Society of In-Vitro Maturation (ISIVM), I extend our sincere
condolences to the Japanese people for the terrible losses and suffering they haverecently incurred. Their bravery and resilience in the face of unspeakable tragedy have been a remarkable
testament to human dignity and endurance.
This meeting marks a special event in the history of reproductive medicine: the presentation of the 2010 Nobel
Prize in Physiology or Medicine to Professor Robert Edwards. On a personal note, Professor Edwards has
played a very special role in my life as a cherished mentor and, along with Howard Jacobs and Stuart Campbell,
as a partner in the London Women's Clinic. I am grateful and privileged to have worked with him and deeply
appreciate the formative role he played in my career.
This well-deserved honor took more than 30 years in coming, but I believe it reflects a recognition of thegrowing importance of reproductive medicine. Following the birth of the first IVF baby through the pioneeringachievements of Professor Edwards, as well as his early work in in-vitro maturation (IVM), ART has advanced
by leaps and bounds over the past three decades.
The advent of IVM heralded a simpler form of treatment without the need for expensive fertility drugs that could
also have unpleasant and potentially dangerous side effects. However, because of its initially poor pregnancy
rates, it has been slow to gain acceptance. But as clinical pregnancy rates/cycle started approach 50% in thebest centres internationally, IVM is rapidly growing in importance as a viable treatment. It is an attractive option
for patients at high risk of ovarian hyperstimulation syndrome, those who have had recurrent unexplained IVF
failures, and those who have had poor or hyper responses to IVF. IVM, followed by oocyte vitrification, also
gives cancer patients, and those with other conditions such as severe SLE, the option of preserving their fertility
without the need for contraindicated gonadotropin stimulation before they undergo oocyte retrieval and oocyte
vitrification. In Montreal, our team has preserved fertility of over 200 women with various medical conditions andin a medical trial achieved a live-birth rate of 20%/cycle through IVM/oocyte vitrification and reported the birth of
the first four healthy babies.
By attending this meeting, you will have the opportunity to learn about the latest advances in IVM and its value
as a complementary treatment to IVF.
As more and more centres provide IVM as an option to their patients, the ISIVM believes it is essential toestablish an international registry to track success rates, pregnancy outcomes, and the health of newborn
babies. This major initiative is important to all ART practitioners and I urge you to join us to make your voiceheard. The benefits of membership are fully explained at our website, www.isivm.com.
I hope you enjoy the Congress and your time in the unique and exciting city of Tokyo.

Seang Lin Tan
President, International Society for In Vitro Maturation

5

WELCOME MESSAGESWELCOME MESSAGE FROM THE CONGRESSPRESIDENTAND THE CONGRESS VICE-PRESIDENT

Dear Colleagues, Ladies and Gentlemen,

Osamu Kato

Hisao Osada

It is our great pleasure to welcome you all to the 16th World Congress on In Vitro Fertilization.
Over the past decade and a half, the World Congress on In Vitro Fertilization has established itself as one of themost significant international gatherings for all doctors and scientists who are working in the field of reproductive
health. Inviting more than 1000 health professionals, including clinicians, scientists, embryologists andprofessionals working in allied fields, the Congress is a vital forum for the exchange of cutting-edge informationin this rapidly advancing field.
We have been preparing to hold this prestigious congress for years and have been really looking forward towelcoming you. In the midst of our preparation, though, the devastating earthquake struck the Tohoku District ofJapan on March 11, 2011, and as a result it nearly became impossible for us to hold this congress. However,
our friends all over the world have extended their kind support to us and our fellow countrymen and women andthanks to you all, we are now able to hold the congress as planned here in Tokyo. For your unflinching support,
we would like to express our deepest gratitude.
Before this trying experience, however, we had been thrilled to receive a wonderful piece of news - thepresentation of the 2010 Nobel Prize in Physiology or Medicine to Professor Robert Edwards, the patron of theInternational Society for In Vitro Fertilization. We ourselves remember well the time he made a visit to our clinicin Tokyo. We are very happy and honoured to have a chance to review and celebrate his achievements in theOpening Ceremony of this Congress.
We have prepared a scientific program for the Congress that focuses on a broad array of fascinating and vital
topics. We are confident that it will prove to be an exciting and rewarding experience for professionals of all kind
involved in our field.
We hope each and every one of you will have an unforgettable, once-in-a-lifetime intellectual and culturalexperience in this great metropolis.

Osamu Kato
Congress President

Hisao Osada
Congress Vice-President

6

COMMITTEESCongress President

Osamu Kato

(Japan)Congress Vice-President

Hisao Osada(Japan)

Pak-Chung Ho
Outi Hovatta
Karl Illmensee
Robert Jansen
Chen Zhi Jiang
Qiao Jie
Steven G. Kaali
Semra Kahraman
Issam Lebbi
Peter Leung
Jin Ho Lim
Jiaen Liu
Alex Lopata
Bruno Lunenfeld
Nalini Mahajan
Khaled Mahmoud
Shin Yong Moon
Huang Guo Ning
Hrishikesh D. Pai
Kuang Yan Ping
Zhou Can Quan
José Remohí
Zev Rosenwaks
Hassan N. Sallam
Marsal Salvina
Joseph Schenker(China)(Sweden)(Greece)(Australia)(China)(China)(USA)(Turkey)(Tunisia)(Canada)(Korea)(China)(Australia)(Israel)(India)(Tunisia)(Korea)(China)(India)(China)(China)(Spain)(USA)(Egypt)(Indonesia)(Israel)International Scientific Committee
Michel Abou Abdallah(Lebanon)
Mohamed Aboulghar(Egypt)Safaa Al-Hasani(Germany)
Gautam Allahbadia(India)Severino Antinori(Italy)Mustafa Bahçeci(Turkey)
Yona Barak(Israel)Artur Bernard(Hungary)Peter Braude(UK)Peter Brinsden(UK)Peter Brockerhoff(Germany)
Luis Arturo Ruvalcaba Castellón (Mexico)
Kwang Yul Cha(Korea)Yoon Seok Chang(Korea)Ri-Cheng Chian(Canada)Patrice Clement(France)
Jacques Cohen(USA)
Aygül Demirol(Turkey)
Klaus Diedrich(Germany)
Jacques Donnez(Belgium)
Jean-Bernard Dubuisson (Switzerland)
Wilfried Feichtinger(Austria)
Dov Feldberg(Israel)Richard Fleming(UK)
René Frydman(France)
Victor Gomel(Canada)Richard Scott
Daniel S. Seidman
Kamala Selvaraj
Gamal A. Serour
Sherman J. Silber
Carlos Simon
Seang Lin Tan
Erol Tavmergen
Tongtis Tongyai
Loy Lan Too
Chii Ruey Tzeng
Filippo Mari Ubaldi
Pierre Vanderzwalmen
Pramuan Virutamasen
Budi Wiweko
P. C. Wong
Wolfgang Würfel
Cao Yun Xia
Frank Yelian
Sergey Yakovenko
Liu Jia Yin
John Yovich
Herbert ZechJohn Zhang
Dominique de Ziegler
Fethi Zhioua(USA)(Israel)(India)(Egypt)(USA)(Spain)(Canada)(Turkey)(Thailand)(Taiwan)(Taiwan)(Italy)(Belgium)(Thailand)(Indonesia)(Singapore)
Germany)(China)(USA)(Russia)(China)(Australia)(Austria)(USA)(France)(Tunisia)(Executive Board members from Japan Society of Fertilization and Implantation)
Yoshihiko Hosoi
Tomohiko Ichikawa
Hiroshi Imai
Minoru Irahara

Osamu Ishihara
Hideharu Kanzaki
Osamu Kato

Yoshiharu Morimoto
Hiroaki Shibahara
Yuji Taketani

Atsushi Tanaka
Osamu Tsutsumi
Kaoru Yanagida

Katsuyuki Kinoshita
Yujiro Kamiguchi
Fumikazu Kotsuji
Koji Koyama
Harumi Kubo
Hirohisa Kurachi
Tsunehisa Makino
Takeshi Maruo
Takashi Minegishi
Kazukiyo Miura

Hideki Mizunuma
Masaru Murai
Yukihiro Nagata
Masahisa Nakamura
Hisashi Narahara
Yoichi Noda
Kahei Sato
Nobuhiko Suganuma
Katsuhiko Takahashi
Toshiyuki Takeshita

Teruhiko Tamaya
Toshinobu Tanaka
Naoki Terakawa
Katsuo Tsubata
Takafumi Utsunomiya
Hideki Yoshida
Yasunori Yoshimura

National Scientific Committee
Tsutomu Araki
Takeshi Aso
Toshio Hata
Kazuhiko Hoshi
Hiroshi Hoshiai
Hiroshi Imai
Noriyuki Inaba
Mutsuo Ishikawa
Bunpei Ishizuka
Mitsutoshi Iwashita

7

COMMITTEES
Yasuhisa Araki
Masaki Inoue
Keiichi Isaka
Hirofumi Kamiya
Toshihiro Kawamura
Toshiro Kubota
Masashige Kuwayama

Koichi Kyono
Yasuhito Michikura
Mineo Morita
Tetsunori Mukaida
Mikio Namiki
Yasushi Odawara
Hidekazu Saito

Makio Shozu
Kou Sueoka
Rikikazu Sugiyama
Yuji Takehara
Toshiyuki Takeshita
Kenichi Tatsumi
Shokichi Teramoto

Kiyotaka Toshimori
Tatsuo Yamamoto
Tetsu Yano
Atsumi Yoshida
Hiroaki Yoshida

Sadao Moridono
Yukio Nakamura

Eimei Sato
Masakuni Suzuki

Shuetsu Suzuki
Takao Yoshida

Hirofumi Kamiya
Yukiko Katagiri
Keiichi KatoToshihiro Kawamura
Koichi Kyono
Tsunekazu Matsumoto
Yasuhito Michikura
Yasuyuki Mio
Yoshiharu Morimoto
Tetsunori Mukaida
Yumi Nagata

Masanori Ochi
Hiroshi Okada
Hisao Osada
Hidekazu Saito
Moritoshi Seki
Masahide Shiotani
Makio Shozu
Kou Sueoka
Rikikazu Sugiyama
Katsuhiko Takahashi
Yuji Takehara

Atsushi Tanaka
Kenichi Tatsumi
Yukihiro Terada
Shokichi Teramoto
Kiyotaka Toshimori
Takafumi Utsunomiya
Naoki Yamashita
Koji Yano
Tetsu Yano
Atsumi Yoshida
Hiroaki Yoshida

National Advisory Committee
Masato Inoue
Akira Iritani
Takahide Mori

Program Committee
Victor Gomel (Canada)
Anis Feki (Switzerland)
Osamu Arakawa
Yoshimasa Asada
Atsushi Azumaguchi
Aisaku Fukuda
Masaki Inoue
Isamu Ishiwata
Hirotsune Kaijima
Hirofumi Kamiya
Hirotsune Kaijima

Board Members of International Society for In Vitro Fertilization (as of May 10, 2010)
President

Yona Barak

Basak Balaban

Moncef Benkhalifa

Svend Lindenberg

Bunpei Ishizuka

Shlomo Mashiach

K.Y. Cha

Lisolette Mettler

Vice-President/Secretary

Ri-Cheng Chian

Luciano Nardo

Victor Gomel

Aygul Demirol

Zev Rosenwaks

Klaus Diedrich

Gerald Schatten

Joshua Dor

Joseph Schenker

Jean-Bernard Dubuisson

Lynette Scott

Alan Handyside

Gamal Serour

Roy Homburg

Ilan Tur-Kaspa

Robert Jansen

Weon Young Son

René Frydman
Past President
Takahide Mori

Treasurer
Seang Lin Tan
Educational Officer
Timur Gürgan
Finance Officer
Barry Cappel
Honorary Patron

Qiao Jie

Jerome Straus

Robert Edwards

Issam Lebbi

Erol Tavmergen

Liaison Tokyo Congress

Milton Leong

Anna Veiga

T.C. Li

Sergey Yakovenko

Osamu Kato &Hisao Osada
Liaison Tunis Workshop
Anis Feki

8

CONGRESS INFORMATION

9CONGRESS INFORMATION

Final Programme & Abstracts
Certificate of Attendance**A Certificate of Attendance is included in the
Final Program & Abstracts (see last page.)

Dates and Venue
Dates: September 10-13, 2011
Venue: Keio Plaza Hotel
2-2-1, Nishi-Shinjuku, Shinjuku-ku, Tokyo
160-8330
Phone: +81-3-3344-0111
www.keioplaza.co.jp

Accompanying person’s registration includes:
Opening & Closing Ceremony
Welcome Reception
Access to the Exhibit Hall
Tokyo City Tour (10th - 12th)
A 3-hour sightseeing tour to visit Meiji Shrine
and Tsukiji Fish Market will be organized forguests free or charge (up to 40 participants pertour):

Contacts
Secretariat of the 16th World Congress on IVF
Secretariat General: Fumihito Aono
c/o Kato Laidies Clinic
7-20-3, Nishi-Shinjuku, Shinjuku-ku, Tokyo 160-0023,
Japan
Fax: +81-3-3366-3908
E-mail: office@isivf2011.org

Sat. Sept. 10: 13:30
Sun. Sept. 11: 9:00 and 13:30
Mon. Sept. 12: 9:00
Please reserve your seat at the “Tokyo City
Tour desk” located in the Reception area on
the forth floor. Reservation will be made on afirst-come, first-served basis.

Registration Office
JTB Global Marketing & Travel Inc., (JTB GMT) has
been appointed as the official registration office forthe congress and will handle registration and hotel
accommodation.

Name Plates
Official Congress name plates will be required for
all Congress functions, scientific sessions and theexhibit area. Individuals who lose their nameplates will be required to pay a fee to obtain apersonalized replacement.

JTB Global Marketing & Travel Inc.
Convention Center (CD102079-041)
2-3-11, Higashi-Shinagawa, Shinagawa-ku, Tokyo
140-8604, Japan
Phone: +81-3-5796-5445
Fax: +81-3-5495-0685
E-mail: ISIVF2011@gmt.jtb.jp

Message Board
A message board will be located near theRegistration Desk. Announcements from thesecretariat or any personal messages will beplaced on the board.

Registration
Registration (Reception) Desk: 4th floor
Opening Hours:
Saturday
September 10
08:00-18:00
SundaySeptember 11
08:00-18:00
MondaySeptember 12
08:00-18:00
Tuesday
September 13
08:00-14:00

Poster Area (Hana BCD, 4F)
The poster area is open at all times on the 4th
floor. Delegates can visit posters at theirconvenience. Poster viewing time is as follows(poster presenters are requested to be at theirposters at those times):

Delegate registration includes:
Opening & Closing Ceremony
Welcome Reception
Access to the Exhibit Hall
Access to all sessions
Delegate Bag

Sunday September 11 10:00-10:30; 13:00-13:20
Monday September 12 10:00-10:30; 13:00-13:20

10

CONGRESS INFORMATION

Commercial Exhibition (Hana BCD, 4F)
Saturday
September 10
08:00-17:00
SundaySeptember 11
08:00-17:00
MondaySeptember 12
08:00-17:00
Tuesday
September 13
08:00-14:30* Coffee and other refreshments are available atthe Exhibition area.
Lunch
Lunch boxes will be provided free of charge forluncheon seminars (you can eat while attending
the lecture during the lunch break). Vouchers forthe lunch boxes will be provided at the dedicateddesk located near the Registration Desk on the
4th floor on a first come first served basis.
Luncheon Seminars are arranged for 12:00-13:00
on Sunday, September 11, and Monday,
September 12 at Concord Ballroom, A, B, and C
on the 5th floor. For topics and speakers, please
see the program.
Coffee and Refreshments
Served in the Commercial Exhibition area at all
times.

11

INSTRUCTIONS for Speakers and Chairpersons

Instructions for Speakers
1. TIME ALLOWANCE FOR PRESENTATION
Plenary Lecture: 30 minutes (25 minutes presentation and 5 minutes discussion)
Symposium: 20 minutes (15 minutes presentation and 5 minutes discussion)
Pre-Congress Workshop: 30 minutes
Free Communication: 15 minutes (10 minutes presentation and 5 minutes discussion)
2. AT THE TIME OF YOUR PRESENTATION- Meet with the session Chairpersons in the session room 15 minutes before the beginning of the session.
- Please be certain that the length of your oral presentation does not exceed the allotted time.
- Chairpersons have been instructed to terminate lectures which exceed their allotted time.
3. EQUIPMENT FOR PRESENTATIONPowerPoint Presentation
Session rooms are provided with one Windows laptop computer on which Microsoft PowerPoint is installed (OS:Windows XP English version; Software: Microsoft PowerPoint 2003/2007/2010) and an LCD projector. The
podium is equipped with a monitor, a mouse and a keyboard. You are requested to bring the data for yourpresentation on a CD-ROM or a USB memory stick, or your own laptop computer.
*Macintosh is not available. If you wish to use a Macintosh PC, please bring your own PC.
1) If you bring data:
You are requested to bring the data for your presentation on a CD-ROM or a USB memory stick to the Speaker
Ready Room (“Kaede” room on the 4th floor) and upload your presentation at least 30 minutes before yoursession begins. Only a CD-ROM in Windows format or USB memory stick is acceptable. The name of the fileshould be labeled with your name. Please also see the <NOTES>.
2) If you bring your own laptop computer:
Windows machines with Windows 98/98SE/2000/ME/XP/VISTA/7 or Macintosh machines with any Mac OS(8.6/9.04-9.2.2/X.1-X.5 and subsequent versions) are acceptable. Please note that Japan operates on 100 voltsfor electrical appliances, with a cycle of 50 Hz in Tokyo.
The plug type in Japan is A with two flat blades. The computer to be used for presentation must be equippedwith a D-sub-15 pin video output. Please make sure that you prepare the necessary converters for yourcomputer so that they meet the requirements stated above. Also, please bring your own AC adaptor.
You are requested to bring your computer and check its connection at the Speaker Ready Room (the “Kaede”
room on the 4th floor) at least 30 minutes before your session begins.
For all computers, check the following settings:-Cancellation of power-saving features: Cancellation of Sleep, Screen Saver, etc.;-Resolution of screen: 1024x768 pixels (XGA) or less.

12

INSTRUCTIONS for Speakers and Chairpersons

In the session room, please have your computer powered on when the presentation prior to your own begins
and hand it over to the operator at the PC desk in the session room, with your presentation file opened. (Youmay be required to restart your computer if the connection to the projector is not successful.) Since there will be
only one connection cable to the projector, the operator will connect it at the beginning of your presentation. The
projector has a standard mini-D-sub 15 pin (three sequences) connector. If your computer has a different
connector, you should bring your own conversion cable.
<NOTES>* When you make a PowerPoint file for your presentation, please make sure that all graphics are embedded in
the presentation file. Fonts should be standard fonts, such as Times New Roman, Arial, or Courier. If
non-standard fonts must be used, they should be embedded in the presentation files. Also, please set up theslide size for “On-screen show”.
* If your presentation includes video/animation, only ones that can be played by “Media Player” are acceptable.
*The name of the file should be labeled with your name.
4. SPEAKER READY ROOM (for slide preview and data check)
The Speaker Ready Room is “Kaede” on the 4th floor. It is open during the following hours:
Saturday
SundayMondayTuesday

September 10
September 11
September 12
September 13

08:00-17:30
08:00-17:30
08:00-17:30
08:00-13:00

Please make sure to visit the room with your data/PC at least 30 minutes before your session begins.
5. NEXT SPEAKER’S SEAT
You are requested to be seated at the Next Speaker‟s seat, located in the left front row 10 minutes before yourpresentation starts. If you are bringing your own PC, please hand it over to the operator at the PC Desk in thesession room.

Instructions for Chairpersons
All session chairpersons are requested to meet with their speakers in the session room 15 minutes before thesession begins.
* Please welcome delegates to the session and introduce each speaker by title, name of all authors, andaffiliation (if any) as listed.
* Any changes to the program should be announced at the beginning of the session.
* An attendant will be present in the session room to assist you with any last minute details.
* Please keep each presentation on schedule. If a presentation is cancelled, do not change the timing of theother presentations.

13

INSTRUCTIONS for Poster Presenters

Instructions for Poster Presenters
Venue: Hana B, C, and D, 4th floor
To provide ample time for all participants to view and discuss poster presentations, each poster will be on
display for the entire duration of the Congress.
Poster presenters are requested to be at their posters during the Poster Viewing Time as follows:
Sunday September 11: 10:00-10:30 and 13:00-13:20
Monday September 12: 10:00-10:30 and 13:00-13:20
Poster presenters are responsible for setting up and removing their posters according to the following schedule:
Setting up: Saturday, September 10, 18:00-20:00 or Sunday, September 11, 8:00-9:00
Removal:Tuesday, September 13, 12:00-15:00<Notes for Poster Presentation>
1. All posters are to be written entirely in English.
2. A presentation number to be placed at the top left of the poster will be provided by the Secretariat. Eachauthor is requested to indicate the “title”, the “authors‟ names” and the “authors‟ affiliations”, at the top rightof the panel within an area measuring 90cm wide by 20 cm high.
3. The usable area for the contents is a space 90cm wide and 160cm high. The layout of thepresentation contents is at the authors‟ discretion.
4. Posters are to be attached to the boards with drawing pins, which will be provided at the Poster Area.
No paste, glue, staples or nails are permitted.
6. Please stay in front of your poster during the Poster Viewing times.
7. The best poster award will be presented in the Closing Ceremony on Tuesday, September 13 (14:3015:00).

No.

Best Abstract Awards
Two abstract awards winners, one for posterand one for oral presentation, will beselected and provided by the International
Society for In Vitro Fertilization. The Winners
will be announced in the Closing Ceremony.

14

FLOOR PLAN OF THE CONGRESS VENUE

Floor Plan of the Congress Venue
Poster & Exhibition Area (Refreshments served)

Hana A

Hana B, C, D

Reception Desk

KAEDE

Vouchers for lunch boxes areprovided here.
Speaker Ready Room (for checking data and PC for presentation)

ConcordBallroom A

ConcordBallroom B

ConcordBallroom C

Eminence HallCOSMOSAZALEA

Secretariat

Conference Room

15

SOCIAL PROGRAM and TRAVEL DESKS

Social Program Official Opening Ceremony and Lecture(Included in registration fee for delegates and registered accompanying persons)Venue: Eminence Hall, 5th floor
Date: Saturday, September 10
Time: 18:30-19:30
Dress Code: Business (e.g. tie and jacket)
 Welcome Reception(Included in registration fee for delegates and registered accompanying persons)Venue: Concord Ballroom ABC, 5th floor
Date: Saturday, September 10
Time: 19:30-21:30
Dress Code: Business (e.g. tie and jacket)
 Congress President’s Dinner(Invited guests only)
Venue: Eminence Hall, 5th floor
Date: Sunday, September 11
Time: 18:30-20:30
Dress Code: Business (e.g. tie and jacket)(An invitation card is included in the delegate bag for the invited guests.)
 Gala Dinner (Buffet)(Requires ticket)
Venue: Concord Ballroom AB, 5th floor
Date: Monday, September 12
Time: 18:30-20:30
Those who wish to take part in the Gala Dinner can purchase tickets at the Registration Desk for
10,000 yen. (An invitation card is included in the delegate bag for the invited guests.)
Dress code: Business/Informal
 ISIVF General Assembly & Closing Ceremony(Included in registration fee for delegates and registered accompanying persons)Venue: Concord Ballroom ABC, 5th floor
Date: Tuesday, September 13
Time: 14:20-15:00
Dress Code: Business/Informal
Travel Desks
 At Narita Airport
A Travel Desk for the Congress will be set up at the Narita Airport to assist your journey to the Congress venue
on September 9 and 10, from 8:00 to 20:00.

16

SOCIAL PROGRAM and TRAVEL DESKS At the Congress Venue (Keio Plaza Hotel)
A Travel Desk operated by Tokyo Metropolitan Government will be opned in the reception area during theCongress. Please contact Travel Desk personnel should you have any questions about your travel
arrangements, sightseeing, etc.

TOKYO CITY TOUR
This tour is specially organized for overseas guests by Tokyo Metropolitan
Government. It is intended for accompanying persons but any delegates can takepart in the tour if seats are available.
Dates:
September 10th (Sat)
September 11th (Sun)September 12th (Mon)13:30-16:30
9:00-12:00 & 13:30-16:30
9:00-12:00

Tour fare: free
Booking:
Please sign up for the tour at “Tokyo City Tour desk” located on the 4th floor.
Max number of participants for each tour is limited up to 40. Reservation will bemade on a first-come, first-served basis.
Tour course:First, visit the tranquil Meiji Shrine, dedicated to the Emperor Meiji, the firstemperor of modern Japan. Then visit Tsukiji Outer Fish Market, where you will
have a chance to try sushi. En route to the hotel, you will enjoy a panoramic viewof the Imperial Palace from the coach.

Please note that the tour course will be subject to change with or without notice.

17

MAP OF THE CONGRESS VENUE

Map of the Congress Venue (Keio Plaza Hotel) and other hotels

6

7
2
4

1

3

5

1. Keio Plaza Hotel Tokyo (Congress Venue)
2. Hyatt Regency Tokyo
3. Hotel Sunroute Plaza Shinjuku
4. Shinjuku Washington Hotel
5. Star Hotel Tokyo
6. Shinjuku New City Hotel
7. Hotel Sunmembers Tokyo Shinjuku

18

ACCESS TO THE CONGRESS VENUE

Access to the Congress Venue (Keio Plaza Hotel) and other hotels
TOKYO INTERNATIONAL AIRPORT(HANEDA)NARITA INTERNATIONAL AIRPORT

We would recommend catching the
Airport Limousine Bus as it takes you
directly to the Keio Plaza Hotel andother hotels nearby in the Shinjuku
Area. The service runs at 20-85 min.
intervals, depending on the time ofday. Please see the timetable. It takes
approx. 85-100 minutes and the fare is3000 yen. Please be sure to choose
the bus bound for your destination
hotel.

JR express train (Narita Express)
takes you to Shinjuku Station inapprox. 90 min at approx. 60 min.
intervals. The fare is 3110 yen.
You can walk or catch a taxi to thehotel from Shinjuku Station.

The Airport Limousine Bus will take you tothe hotels in the Shinjuku area, including
the Keio Plaza Hotel, Shinjuku Washington
Hotel & Hyatt Regency Hotel.The fare is1200 yen and it takes approx. 60 min.
Please be sure to choose the bus bound foryour destination hotel.

JR SHINJUKU STATION

10 min. walk

3 min. walk
9 min. walk

SHINJUKUWASHINGTON
HOTEL

HYATT
REGENCY
TOKYO

5 min. walk

15 min. walk
3 min. walk

SUNROUTEPLAZASHINJUKU

STAR
HOTEL
TOKYO

10
7 min.
min.walkwalk

4 min. walk

15 min. walk

SHINJUKUNEW CITY
HOTEL

HOTEL
SUNMEMBERSTOKYO
SHINJUKU

7 min. walk

7 min. walk

10 min. walk

KEIO PLAZA HOTEL TOKYO (CONGRESS VENUE)

For details of the Airport Limousine Bus service from Narita Airport and Haneda Airport to Keio Plaza
Hotel, please see http://www.limousinebus.co.jp/en/platform_searches/index/2/18

19

ACCESS TO THE CONGRESS VENUE

Airport Limousine Bus Timetable: from Narita Airport to Keio Plaza Hotel
Narita Airport
Terminal 2

07:00

07:40

08:30

09:20

10:20

11:00

11:40

12:20

13:00

13:25

Narita Airport
Terminal 1
South Wing

07:05

07:45

08:35

09:25

10:25

11:05

11:45

12:25

13:05

13:30

Narita Airport
Terminal 1
North Wing

07:10

07:50

08:40

09:30

10:30

11:10

11:50

12:30

13:10

13:35

Keio Plaza Hotel

09:05

09:45

10:35

11:25

12:25

13:05

13:45

14:25

15:05

15:30

Narita Airport
Terminal 2

13:45

14:30

15:00

15:45

16:30

17:00

17:45

18:15

18:45

19:40

Narita Airport
Terminal 1
South Wing

13:50

14:35

15:05

15:50

16:35

17:05

17:50

18:20

18:50

19:45

Narita Airport
Terminal 1
North Wing

13:55

14:40

15:10

15:55

16:40

17:10

17:55

18:25

18:55

19:50

Keio Plaza Hotel

15:50

16:35

17:05

17:50

18:35

19:05

19:45

20:15

20:40

21:30

Narita Airport Terminal 2

20:10

21:00

21:40

22:35

Narita Airport Terminal 1 South Wing

20:15

21:05

21:45

22:40

Narita Airport Terminal 1 North Wing

20:20

21:10

21:50

22:45

Keio Plaza Hotel

22:00

22:50

23:25

00:15

Airport Limousine Bus Timetable: from Keio Plaza Hotel to Narita Airport
Keio Plaza Hotel

05:50

06:50

07:50

08:45

09:50

10:50

11:50

12:50

13:50

14:50

Narita Airport
Terminal 2

07:25

08:25

09:40

10:50

11:50

12:50

13:45

14:40

15:40

16:40

Narita Airport
Terminal 1

07:30

08:30

09:45

10:55

11:55

12:55

13:50

14:45

15:45

16:45

Keio Plaza Hotel

15:50

16:45

17:45

Narita Airport
Terminal 2

17:50

18:50

19:50

Narita Airport
Terminal 1

17:55

18:55

19:55

20

LOCAL INFORMATION

General information about Japan
Population:
Capital:Currency:

Around 128 millionTokyo
Yen (usually indicated as ¥)The yen comes in the following denominations: Coins: 1, 5, 10,50, 100, 500 yen
Bank notes: 1000, 2000 (very rare), 5000, 10000 yen.
Climate:Average temperature in September: 23 degrees C.
Units of measure: Metric system
Electricity:
100V, with 50 Hz in east Japan and 60 Hz in west Japan
Time Zone:
GMT + 9Passports & Visa: If you are a citizen of one of the over 50 countries with which
Japan has a "general visa exemption arrangement", you need only a valid passport to enter Japanas a "temporary visitor". Otherwise, you need to obtain a visa before entering the country. Temporary
visitors from most countries are allowed to stay for up to 90 days.
Consumption Tax: Paid by consumers when they purchase goods and services. The current rate is 5%. Shops andother service providers are required to include the consumption tax in the prices shown.
Tipping:
Basically, it is not a custom to give tips, except in high class ryokan (traditional Japanese inn.)

About Tokyo
Tokyo, a sprawling metropolis with a population of nearly 13 million people, is one of the world‟s great cities. Originally asmall castle town in the 16th century, by the 18th century it was the centre of the Shogun‟s power and already one of thelargest cities in the world. Since the middle of the 19th century, it has been the capital and the financial and cultural hub ofmodern Japan.
With this history behind it, Tokyo today is a fascinating blend of the traditional, the modern and the hyper-modern, afascinating blend of East and West, and yet at the same time a uniquely Japanese city. Tokyo really is a place that has
something for everyone – a place traditional temples and shrines stand cheek by jowl with towering skyscrapers, andsmall traditional bars and shops can be found just behind some of the most exciting and innovative architecture to befound anywhere in the globe.
With its super-efficient transport system and its extremely low rates of crime, Tokyo is a city where one can enjoy the bestthat the world can offer, whether it is in terms of art and culture, cuisine, the latest hi-tech gadgetry, financial wizardry, or
human warmth and friendliness. Whatever your interests, Tokyo can offer you an experience that you will never forget.
For more information, please visit: http://www.japan-guide.com

Accompanying Person’s TourWhy not join the 3-hour, free sightseeing tour to visit Meiji Shrine and Tsukiji Fish Market? The tour starts
in front of the Keio Plaza Hotel at the following hours: 9:00 for September 11 and 12; 13:30 for September
10 and 11. Please reserve your seat at the Accompanying Person‟s Tour Desk located next to the
Reception Desk on the 4th floor of Keio Plaza Hotel. Seats are provided on a first come first served basis.

21

LOCAL INFORMATION

Traditional Sightseeing Spots in Tokyo
The Imperial Palace
The current Imperial Palace (Kokyo) is located on the former site of Edo Castle, alarge park area surrounded by moats and massive stone walls in the center of Tokyo,
a short walk from Tokyo Station. It is the residence of Japan's Imperial Family. The
Imperial Palace East Gardens are open to the public throughout the year except on
Mondays, Fridays and special occasions.
Asakusa
Asakusa is the center of Tokyo's “shitamachi”, lit. "low city", one of the districts ofTokyo which have preserved a certain atmosphere of the old city. Asakusa„s main
attraction is Sensoji, a very popular Buddhist temple, built in the 7th century.
The temple is approached via the Nakamise, a shopping street that has been
providing temple visitors with a variety of traditional, local snacks and tourist
souvenirs for centuries.

Sightseeing Spots in Shinjuku
Shinjuku is one of the 23 wards of Tokyo, but the name commonly refers just to thelarge entertainment, business and shopping area around Shinjuku Station. Handlingmore than two million passengers each day, Shinjuku Station is Japan„s busiest
railway station, served by six railway companies and about a dozen railway andsubway lines, including the JR Yamanote Line.
Tokyo Metropolitan Government Office (Tocho)
The 243 meter tall twin towers and surrounding buildings contain the offices and theassembly hall of the metropolitan government of Tokyo, as well as observatories on
the 45th floor of each tower. The view from the southern tower is considered the
best.
Open daily 9:30 to 23:00 (south observatory until 17:30), except December 29-31,
January 2-3 and occasional inspection days. Furthermore, the north observatory is
closed on the 2nd and 4th Monday and the south observatory on the 1st and 3rd Tuesday of each month, except if apublic holiday falls on the closure day, in which case the observatory is closed the following day. Admission is free.
Shinjuku Gyoen
Shinjuku Gyoen is one of Tokyo's largest and most pleasant parks andbest cherry blossom viewing spots. It was opened to the public in 1949,
after it had served as a garden for the Imperial Family since 1903. Openfrom 9:00 to 16:30. Closed on Mondays (Tuesday when Monday is anational holiday) from December 29 to January 3. There are no closuredays during the cherry blossom season (late March to late April) and theChrysanthemum Exhibition (first half of November). Admission is 200 Yen.

22

LOCAL INFORMATION

Museums (Please note that entries are permitted until 30 minutes before the closing.)
Tokyo National Museum / Ueno
The Tokyo National Museum collects, houses, and displays acomprehensive collection of art works and antiquities from Japan aswell as other Asian countries.
Access: 10 mins from JR Ueno Station
Opening Hours: Tue.-Thu.: 9:30-17:00, Fri.: 9:30-20:00
Sat. & Sun.: 9:30-18:00 Closed on Mondays except Sept. 15

National Museum of Nature and Science / Ueno
The Museum boasts one of the richest histories of any museum in Japan.
It is Japan's only nationally administered museum, and one of the world's
central institutes for research in natural history and the history of scienceand technology.
Access: 5 mins walk from JR Ueno Station
Opening Hours: 9:00-17:00, Closed on Sept. 5, 12, 20 & 26.

National Museum of Western Art / Ueno
The NMWA was established in 1959 around the core Matsukata
Collection as Japan's museum specializing in Western art. The galleries
feature pre-18th century paintings including those by Ritzos, Van Cleve,
Veronese, Rubens, Van Ruysdael, and Ribera, 19th to early 20th century
French paintings including works by Delacroix, Courbet, Manet, Renoir,
Monet, Van Gogh, Gauguin, and Moreau and works by the next generation ofartists, such as Marquet, Picasso, Soutin, Ernst, Miro, Dubuffet and Pollock.
Access: 1 min. walk from JR Ueno Station
Opening Hours: Normally 9:30-17:30, Closed on Mondays

Mori Art Museum / Roppongi
ROPPONGI HILLS MORI TOWER (53F)The Musium takes the lead in introducing the newest art from Asia and otherregions of the world. Key enphasis is placed on the concepts of being“Contemporary” and “International”.
Access (To Roppongi Hills): 4 min. walk from Exit 3, Roppongi Station
on Toei Oedo Subway Line
Opening Hours: Everyday except Tuesdays: 10:00-22:00 (Tuesday until 17:00)

The National Art Center, Tokyo / Roppongi
The National Art Center, Tokyo is a unique and innovative art
exhibition facility: Instead of maintaining a permanentcollection, it makes the most of a total of 14,000 square meters of exhibition
space, one of the largest in Japan, and focuses on serving as a venue forvarious art exhibitions.
Access: 4-minute walk from Exit 7, Roppongi Station on Toei Oedo Subway line
Opening Hours: Mon, Wed, Thu, Sat, and Sundays 10:00-18:00,
Friday 10:00-20:00, Closed on Tuesdays

23LOCAL INFORMATIONThe Edo-Tokyo Museum / Ryogoku
The Edo-Tokyo Museum was founded in 1993, as a facility to preserve thehistorical heritage of Edo -Tokyo. In the Permanent Exhibition area,
there can be found original and replicated exhibits, as well as large-scale models,faithful representations of their originals. Also at t the Special exhibition gallery,visitors can enjoy selected exhibits on subjects related to Tokyo‟s history andculture, scheduled several times a year. The building was modeled after anelevated-floor type warehouse, the height of which is approximately the same asthat of Edo Castle tower.
Access: 3-minute walk from Ryogoku Station (West Exit) on JR Sobu Line
Opening Hours: 9:30-17:30 (Saturday until 19:30), Closed on Mondays

Away from Tokyo
Hakone (2 hours from Shinjuku)
Hakone is part of the Fuji-Hakone-Izu National Park, less than 100 kilometersfrom Tokyo. Famous for hot springs, outdoor activities, natural beauty and theview of nearby Mt. Fuji, Hakone is one of the most popular destinations among
Japanese and international tourists looking for a break from Tokyo.
Mount Fuji (2.5 hours from Shinjuku)
Mount Fuji (Fujisan) is the Japan's highest mountain. It is not surprising that thenearly perfectly shaped volcano has been worshipped as a sacred mountain andloved by both artists and common people.
The Hakone Open Air Museum (2.5 hours from Shinjuku)
Access: JR Odawara Stn. (or Odakyu line Odawara Stn. ) →Hakone Tozan
Railway Chokoku-no-mori Stn. (2min walk)Open Hours: 9:00-17:00, Open year-round
Admission Fee: Adults 1600 yen, Senior citizens(65 and above) and university
and high school students 1100yen, middle and elementary school students 800yen
Kamakura (1 hour from Shinjuku)
Kamakura is a very popular tourist destination. Sometimes called the Kyoto ofEastern Japan, Kamakura offers numerous temples, shrines and other historicalmonuments. In addition, Kamakura's sand beaches attract large crowds during
the summer months.
Nikko (2 hours from Shinjuku)
Nikko is a small city at the entrance to Nikko National Park. It is most famous for
the Toshogu, Japan's most lavishly decorated shrine complex and mausoleum of
Tokugawa Ieyasu, the founder of the Tokugawa shogunate.
Nikko has been a center of Shinto and Buddhist mountain worship for many
centuries, and Nikko National Park continues to offer scenic, mountainous
landscapes, lakes, waterfalls, hot springs, wild monkeys and hiking trails.

24

LOCAL INFORMATION

Kyoto (2 hours and 40 minutes from Tokyo Station by Bullet Train)
Kyoto was Japan's capital and the Emperor's residence from 794 until 1868. It is
now the country's seventh largest city with a population of 1.4 million people and amodern face. Over the centuries, Kyoto was destroyed by many wars and fires,but due to its historic value, the city was dropped from the list of target cities forthe atomic bomb and spared from air raids during World War II. Countless
temples, shrines and other historically priceless structures survive in the city
today.

Nara (1 hour from Kyoto)
Japan's first permanent capital was established in the year 710 at Heijo, the city
now known as Nara. As the influence and political ambitions of the city's powerful
Buddhist monasteries grew to become a serious threat to the government, thecapital was moved to Nagaoka in 784. Nara is located in the Kinai plain, less than
one hour from Kyoto and Osaka. Due to its past as the first permanent capital, it
remains full of historic treasures, including some of Japan's oldest Buddhist
temples.

Taxi
Taxi fares typically start from 710 yen for the first two kilometers and increase by roughly 90 yen for every additional 288
meters traveled. The cost also increases when the taxi is not moving for a prolonged time. Late in the evening, rates areraised by 20-30 percent. A plate on the dashboard in the lower corner of the windshield indicates whether a taxi is vacantor not. A red plate indicates that the taxi is vacant, while a green plate indicates the opposite. When you board a taxi, notethat the vehicle's left rear door is opened and closed remotely by the driver. You are not supposed to open or close it by
yourself. Furthermore, you are not supposed to tip taxi drivers, as the service is included in the price.
If you do not speak Japanese, or your destination is not a well known place, it is recommended that you give your driverthe precise address of your destination on a piece of paper or, even better, point it out on a map, since the Japanese
address system can be confusing even to local taxi drivers.

Food
Japanese cuisine offers a great variety of dishes and regional specialties. Some of the most popular Japanese andJapanized dishes are listed below.
Sushi
Sushi can be defined as a dish which contains sushi rice, cooked rice that is prepared withsushi vinegar. There are various kinds of sushi dishes.
TempuraTempura is seafood, vegetables, mushrooms and other pieces of food coated with tempura batterand deep fried. Tempura was introduced to Japan by the Portuguese in the 16th century, but has
become one of Japan's most famous dishes internationally.
Sukiyaki
A nabe dish prepared with thinly sliced meat, vegetables, mushrooms, tofu and shirataki (konyaku
noodles). The pieces of food are dipped into a raw egg before eaten.

25

LOCAL INFORMATION

Shabu-Shabu
Shabu-shabu is Japanese style meat fondue. Thinly sliced meat, along with vegetables,
mushrooms and tofu is dipped into a hot soup and then into ponzu vinegar or a sesame sauce
before being eaten.
Soba
Soba noodles are native Japanese noodles made of buckwheat flour or a mixture of buckwheat
and wheat flour. Soba are about as thick as spaghetti. They can be served cold or hot and withvarious toppings.
Ramen
Ramen are Chinese style noodles prepared in a soup with various toppings. Ramen is one of themany popular dishes that were originally introduced from China but have become completely
Japanized over time.
Yakitori
Yakitori are grilled chicken pieces on skewers. Most parts of the chicken can be used for yakitori.

26

AGENDA-at-a-Glance
Concord A (5F)

Saturday, September 10, 2011
Concord B (5F)

Concord C (5F)

Hana A (4F)A
9:00

HANA BCD (4F)9:00-12:00
Pre-Congress Workshop[Vitrification]

Chair: Stanley Leibo (USA)
Sherman Silber (USA)(1) Lectures
Stanley Leibo (USA)

10:00

Noriko Kagawa (Japan)
Sherman Silber (USA)(2) Live demonstration
Masashige Kuwayama(Japan)the29th Annual Meeting
11:00

of Japan Society of Fertilization
and Implantation

12:00

13:00

14:00

14:00-17:00
Pre-Congress Workshop[PGD]

Chair: Naoki Aoyama (Japan)
Ilan Tur-Kaspa (USA)(1) Lectures
Kou Sueoka (Japan)
Suguru Sato (Japan)

15:00

Ilan Tur-Kaspa (USA)(2) Live demonstration
Naoki Aoyama (Japan)

16:00

17:00

18:00

18:30-19:30 Opening Ceremony and Lecture (Venue: Eminence Hall ・5F)
Speaker: Victor Gomel (Canada)
19:30-21:30 Welcome Reception (Venue: Concord ABC ・5F)

27

8:30-17:30
Exhibits

AGENDA-at-a-Glance

Sunday, September 11, 2011

Concord A (5F)

9:00

10:00

Concord C (5F)

HANA A (4F)

10:00-10:30
Coffee Break & Poster Viewing
10:30-11:50
C-1 Management of PCO patients (1)

11:00

Concord B (5F)

Chair: Timur Gürgan (Turkey)
Chair: Makio Shozu (Japan)
1 Takahide Mori (Japan)
2 Rogerio A. Lobo (USA)
3 Muchsin Jaffar (Indonesia)
4 Khaled Mahmoud (Tunisia)

10:30-11:50
C-4 Treatment of benign tumor prior to
IVF-ET (1)Chair: Zion Ben-Rafael (Israel)
Chair: Hassan Sallam (Egypt)
1 Hassan Sallam (Egypt)
2 Issam Lebbi (Tunisia)
3 Hiroshi Nabeshima (Japan)
4 Victor Gomel (Canada)

8:30-18:00

10:30-11:50

10:30-12:00
Oral Communications

C-8 ART-cryopreservation: Is oocyte
cryopreservation a revolution in IVF?
Chair: Stanley Leibo (USA)
Chair: Masashige Kuwayama(Japan)1 Luis Arturo Ruvalcaba
Castellón (Mexico)
2 Masashige Kuwayama(Japan)3 Safaa Al Hasani (Germany)
4 Stanley Leibo (USA)

12:00

12:00-13:00
12:00-13:00
12:00-13:00
Luncheon Seminar 1
Luncheon Seminar 2
Luncheon Seminar 3
GnRH antagonists in ovarian stimulation Clinical aspects of GnRH analogue use in Potential of Piezo-Assisted
for IVF and ICSI from practical
Assisted reproduction
Micromanipulation
perspective
Chair: Osamu Ishihara (Japan)
Chair: Fumihito Aono (Japan)
Chair:
Speaker:
Speaker:
Takafumi Utsunomiya (Japan)
Naoaki Kuji (Japan)
Teruhiko Wakayama (Japan)
Speaker: Atsushi Tanaka (Japan)
Hiroshi Morita (Japan)
Sponsored by
Akiko Yabuuchi (Japan)
Sponsored by MSD K.K.
Mochida Pharmaceutical Co., Ltd.
Sponsored by Prime Tech Ltd.
13:00
13:00-13:20
Coffee Break & Poster Viewing
13:20-15:00
C-2 Management of PCO Patients (2)

14:00

15:00

Chair: Rogerio A. Lobo (USA)
Chair: Khaled Mahmoud (Tunisia)
1 Makio Shozu (Japan)
2 Chii-Ruey Tzeng (Taiwan)
3 Toshiro Kubota (Japan)
4 Timur Gürgan (Turkey)
5 Hiroaki Shibahara (Japan)

15:00-15:20
Coffee BreakSponsored by
Mochida Pharmaceutical Co., Ltd.
15:00-15:20
Coffee Break
15:20-16:40
C-6 Oncofertility - Fertility preservation in
women with malignant disease (1)1 Bruno Lunenfeld (Israel)
2 Baris Ata (Canada)
3 Maximilian Murtinger(Austria)
4 Pak-Chung Ho (Hong Kong)

C-3 Controlled ovarian stimulation (2)18:00

Chair: Victor Gomel (Canada)
Chair: Issam Lebbi (Tunisia)
1 Jean-Bernard Dubuisson(Switzerland)
2 Yuji Hiramatsu (Japan)
3 Zion Ben-Rafael (Israel)
4 Hisao Osada (Japan)
5 Michael DeRosa (USA)

15:20-16:40

Sponsored by MSD K.K.
16:40-18:00

17:00

13:20-15:00
C-5 Treatment of uterine myoma or
adenomyosis prior to IVF-ET (2)C-3 Controlled ovarian stimulation (1)Chair: Bruno Lunenfeld (Israel)
Chair: Milton Leong (Hong Kong)

16:00

HANA BCD (4F)

8:20-8:30 Opening Remarks
8:30-10:00
Plenary Lectures
1 Chair: Victor Gomel (Canada)
Speaker: Osamu Kato (Japan)
Speaker: John Zhang (USA)
2 Chair: Sherman Silber (USA)
Speaker: Zsolt Peter Nagy (USA)
3 Chair: Takahide Mori (Japan)
Speaker: Shin Yong Moon (Korea)

8:30

Chair: Svend Lindenberg(Denmark)
Chair: Minoru Irahara (Japan)
5 John Zhang (USA)
6 Svend Lindenberg (Denmark)7 Hassan Sallam (Egypt)
8 Zion Ben-Rafael (Israel)

Chair: Claus Yding Andersen(Denmark)
Chair: Takeshi Maruo (Japan)
1 Rufino Garcia-Otero Reina(Spain)2 Werner Lichtenegger(Germany)
3 Takuma Fujii (Japan)
4 Masaaki Ando (Japan)
16:40-18:00
C-7 Oncofertility - Fertility preservation in
women with malignant disease (2)Chair: Klaus Diedrich (Germany)
Chair: Kazunori Ochiai (Japan)
1 Shokichi Teramoto (Japan)
2 Claus Yding Andersen(Denmark)
3 Liong Tulusan (Germany)
4 Bruno Lunenfeld (Israel)

13:20-14:40

13:20-14:40

C-9 Children follow-up in ART (1)C-12 Laboratory advances in IVF (2)Chair: Fethi Zhioua (Tunisia)
Chair: Osamu Ishihara (Japan)
1 Jia-Yin Liu (China)
2 Ilan Tur-Kaspa (USA)
3 Yukiko Katagiri (Japan)
4 Dov Feldberg (Israel)

Chair: Yoshihiko Hosoi (Japan)
Chair: Zsolt Peter Nagy (USA)
1 Tsunehisa Makino (Japan)
2 Tae-Ki Yoon (Korea)
3 Pierre Vanderzwalmen(Belgium)
4 Gábor Vajta (Australia)

14:40-15:00
Coffee Break

14:40-15:00
Coffee Break

15:00-16:20

15:00-18:00
Oral Communications

C-10 Children follow-up in ART (2)Chair: Hideharu Kanzaki (Japan)
Chair: Dov Feldberg (Israel)
1 Peter Brockerhoff(Germany)
2 Fethi Zhioua (Tunisia)
3 Ri-Cheng Chian (Canada)
4 Tetsunori Mukaida (Japan)

16:20-17:40
C-11 Laboratory advances in IVF (1)Chair: Pierre Vanderzwalmen(Belgium)
Chair: Gábor Vajta (Australia)
1 Artur Bernard (Hungary)
2 Guoning Huang (China)
3 Nicolas Zech (Austria)
4 Zsolt Peter Nagy (USA)Sponsored by MSD K.K.

18:30-20:30 President’s Dinner (Venue: Eminence Hall ・5F)

28

Exhibits and Poster Viewing

AGENDA-at-a-Glance

Monday, September 12, 2011

Concord A (5F)
8:30

Concord B (5F)

Concord C (5F)

10:00

10:00-10:30
Coffee Break & Poster Viewing
10:30-11:50

12:00

10:30-12:10
C-16 Embryo development andC-13 Controlled ovarian stimulation (3)competency from activation to
implantation
Chair: Herbert Zech (Austria)
Chair: Osamu Tsutsumi (Japan)
Chair: Jin-Ho Lim (Korea)
1 Bill Yee (USA)
Chair: Atsushi Azumaguchi (Japan)
2 Milton Leong (Hong Kong)
1 Nelly A. Frydman (France)
3 Johan Smitz (Belgium)
2 Moncef Benkhalifa (France)
4 Herbert Zech (Austria)
3 Atsushi Azumaguchi (Japan)
4 Svend Lindenberg (Denmark)
5 Peter Kovacs (Hungary)

12:00-13:00
Luncheon Seminar 4

14:00

15:00

Chair:
Yoshiharu Morimoto (Japan)
Speaker:
Budi Wiweko (Indonesia)
Sponsored by Medical ＆ Biological
Laboratories Co.,Ltd.

14:20-15:20
C-14 Advances in basic research andlaboratory techniques in IVM (2)Chair: Jie Qiao (China)
Chair: Hiroaki Yoshida (Japan)
4 Ri-Cheng Chian (Canada)
5 Yoshiharu Morimoto (Japan)
6 Hai Ying Chen (Canada)

14:20-15:20
C-18 Update on male infertility - IMSI/ICSI/MD-TESE/PGD
Chair: Sherman Silber (USA)
Chair: Hiroshi Okada (Japan)
1 Sherman Silber (USA)
2 Atsumi Yoshida (Japan)
3 Pierre Vanderzwalmen(Belgium)

15:40-17:20
16:00

17:00

Sponsored by
Tosoh Corporation
13:00-13:20
Coffee Break & Poster Viewing
13:20-14:20
C-17 ART-Gamete manipulation
Chair: Moncef Benkhalifa (France)
Chair: Atsushi Tanaka (Japan)
1 Akiko Yabuuchi (Japan)
2 Moncef Benkhalifa (France)
3 Atsushi Tanaka (Japan)

C-15 Clinical outcomes; toward a more
successful IVM
Chair: Johan Smitz (Belgium)
Chair: Aisaku Fukuda (Japan)
1 Jie Qiao (China)
2 Shu Hashimoto (Japan)
3 Jin-Ho Lim (Korea)
4 Aisaku Fukuda (Japan)
5 Svend Lindenberg (Denmark)8:30-18:00
10:30-12:00
Oral Communications

Chair:
Yuji Takehara (Japan)
Speaker:John Zhang (USA)

13:20-14:20
C-14 Advances in basic research andlaboratory techniques in IVM (1)Chair: Yoshiharu Morimoto (Japan)
Chair: Ri-Cheng Chian (Canada)
1 Nelly A. Frydman (France)
2 Hiroaki Yoshida (Japan)
3 Johan Smitz (Belgium)

15:20-15:40
Coffee Break

10:30-11:50
C-19 AMH: Is it essential for assessing
ovarian reserve
Chair: Bunpei Ishizuka (Japan)
Chair: Hsin-Fu Chen (Taiwan)
1 Yoshimasa Asada (Japan)
2 Budi Wiweko (Indonesia)
3 Bunpei Ishizuka (Japan)
4 Hsin-Fu Chen (Taiwan)

12:00-13:00
Luncheon Seminar 5
Fertility tourism in the U.S.

The role of AMH in female reproduction

13:00

HANA BCD (4F)

8:30-10:00
Plenary Lectures
4 Chair: Atsushi Tanaka (Japan)
Speaker: Alan Handyside (UK)
5 Chair: Chii-Ruey Tzeng (Taiwan)
Speaker: Sherman Silber (USA)
6 Chair: Shuetsu Suzuki (Japan)
Speaker: Victor Gomel (Canada)

9:00

11:00

HANA A (4F)

13:20-14:40
C-20 The role of ART in international
development (1)Chair: Willem Ombelet (Belgium)
Chair: Sheryl Vanderpoel(Switzerland)
1 Shangwei Li (China)
2 Gautam Allahbadia (India)
3 Daniel S. Seidman (Israel)
4 Oriol Coll (Spain)

14:40-15:40
C-20 The role of ART in international
development (2)Chair: Daniel Seidman (Israel)
Chair: Tetsunori Mukaida (Japan)
5 Claus Peter Janisch (Germany)
6 Sheryl Vanderpoel (Switzerland)
7 Willem Ombelet (Belgium)

15:20-15:40
Coffee Break

13:20-15:00
APART Session: MSCs - From basic
research to clinical applications
Chair: Yuji Takehara (Japan)
Chair: Byung-Rok Do (Korea)
1 Byung-Rok Do (Korea)
2 Ken Nakama (Japan)
3 Noriko Kagawa (Japan)
4 Rie Yamadera (Japan)
5 Yuji Takehara (Japan)

15:00-15:20
Coffee Break
15:20-17:50
Oral Communications

15:40-16:00
Coffee Break

15:40-17:40
ISF Session by the Israeli Society of
Fertility
Chair: Arye Hourwitz (Israel)
Chair: Martha Dirnfeld (Israel)
1 Yaron Rabinovici (Israel)
2 Arye Hourwitz (Israel)
3 Martha Dirnfeld (Israel)
4 Daniel Seidman (Israel)
5 Ariel Revel (Israel)
6 Shevach Friedler (Israel)

16:00-18:00
STGO Session by the Tunisian Society
of Gynecology and Obstetrics
Challenging issues in ART (1)Chair: Hedi Khairi
Chair: Anis Feki
Chair: Said Lazrak
1 Youssef Boutaleb (Morocco)
2 René Frydman (France)
3 Amina Oumziane (Algeria)
Challenging issue in ART (2)Chair: Mahmoud Kharouf
Chair: Moise Fiadjoe
Chair: Nabil Ben Zineb
1 Khaled Terras (Tunisia)
2 Moncef Benkhalifa (France)
3 Merdassi Ghaya (Tunisia)

18:00
18:30-20:30 Gala Dinner (Venue: Concord AB・5F)

29

Exhibits and Poster Viewing

AGENDA-at-a-Glance

Tuesday, September 13, 2011

Concord A (5F)
8:30

10:00-10:30
Coffee Break

10:00-10:30
Coffee Break

10:30-11:50
10:30-11:50
C-21 Repeated implantation failure: C-22 Pre-implantation genetic
how to improve implantation in ART screening: an update

11:00

Concord C (5F)

HANA BCD (4F)

Chair: Kouichi Takakuwa(Japan)Chair: Yael Gonen (Israel)
1 Yael Gonen (Israel)
2 Kouichi Takakuwa (Japan)
3 Hiroshi Fujiwara (Japan)
4 Moritoshi Seki (Japan)

Chair: Hidekazu Saito (Japan)
Chair: Jiaen Liu (China)
1 Atsushi Tanaka (Japan)
2 Wilfried Feichtinger (Austria)
3 Kou Sueoka (Japan)
4 Alan Handyside (UK)10:30-12:10
C-23 Stem cell and fertility
preservation: Where are we at withstem cells?
Chair: Kwang Yul Cha (Korea)
Chair: Oktay Kadayifci (Turkey)
1 Hidenori Akutsu (Japan)
2 Anis Feki (Switzerland)
3 Hsin-Fu Chen (Taiwan)
4 Kwang Yul Cha (Korea)
5 Takehiko Ogawa (Japan)

12:10-13:10
Luncheon Seminar 6
Global inequity: Access to Assisted
Reproductive Technologies
Chair:
Osamu Ishihara (Japan)
Speaker:
Sheryl Vanderpoel (Switzerland)
13:00-13:30
Coffee Break

8:00-14:30
Exhibits andPoster Viewing

10:00-10:30
Coffee Break

12:00

13:00

HANA A (4F)

8:30-10:00
Plenary Lectures
7 Chair: Yoon-Seok Chang (Korea)
Speaker: Klaus Diedrich (Germany)
8 Chair: Hiroshi Okada (Japan)
Speaker: Gianpiero D. Palermo (USA)
9 Chair: Yoshiharu Morimoto (Japan)
Speaker: Seang Lin Tan (Canada)

9:00

10:00

Concord B (5F)

13:00-13:30
Coffee Break

13:10-13:30
Coffee Break

13:30-14:30
Special Guest Lecture
Chair:
Akira Iritani (Japan)
Speaker: Shinya Yamanaka (Japan)

14:00

14:30-15:00
ISIVF General Assembly & Closing Ceremony
15:00

30

10:30-12:00
Oral Communications

ANNOUNCEMENT of The 17th World Congress on In Vitro Fertilization, Tunis, Tunisia, 2013

Tunis

31

SCIENTIFIC PROGRAM

32

SCIENTIFIC PROGRAM

1. Special Guest Lecture:
Tuesday, September 13

13:30～14:30 (Concord Ballroom ABC)
Chairperson: Akira Iritani (Japan)

13:30～14:30

Induction of pluripotency by defined factors- Shinya Yamanaka (Japan)

2. Opening Ceremony, Opening Lecture and Welcome Reception:
Opening Ceremony: Saturday, September 10 18:30～19:30 (Eminence Hall, 5F)
18:30～19:00
Opening Ceremony
19:00～19:30
Opening
A tribute to Robert Edwards - the pioneer of human IVF
Lecture-Victor Gomel (Canada)
Welcome Reception: Saturday, September 10

19:30～21:30 (Concord Ballroom ABC, 5F)

3. Plenary Lectures:
Sunday, September 11 8:30～10:00(Concord Ballroom ABC)
8:30～8:50
Plenary
Chairperson: Victor Gomel (Canada)
Lecture -1 (1)Advances in natural and mild stimulation cycle IVF- progress over the last 15 years-Osamu Kato (Japan)
8:50～9:00

9:00～9:30

9:30～10:00

Plenary
Lecture -1 (2)Plenary
Lecture-2

Plenary
Lecture -3

Chairperson: Victor Gomel (Canada)

Clomid® ovarian stimulation for IVF and frozen ET-John Zhang (USA)
Chairperson: Sherman Silber (USA)Non-invasive assessment of embryo quality-Zsolt Peter Nagy (USA)
Chairperson: Takahide Mori (Japan)

The prospect of embryonic stem cell research-Shin Yong Moon (Korea)

Monday, September 12 8:30～10:00 (Concord Ballroom ABC)
8:30～9:00
Plenary
Chairperson: Atsushi Tanaka (Japan)
Lecture-4
Pre-implantation genetic diagnosis: an update-Alan Handyside (UK)9:00～9:30

9:30～10:00

Plenary
Lecture -5

Plenary
Lecture -6

Chairperson: Chii-Ruey Tzeng (Taiwan)

Preservation of fertility-Sherman Silber (USA)
Chairperson: Shuetsu Suzuki (Japan)

The role of reproductive surgery in the era of IVF&ART-Victor Gomel (Canada)33

SCIENTIFIC PROGRAM

Tuesday, September 13 8:30～10:00 (Concord Ballroom ABC)
Plenary
8:30～9:00
Chairperson: Yoon-Seok Chang (Korea)
Lecture -7
Recent advance of controlled ovarian stimulation-Klaus Diedrich (Germany)
9:00～9:30

9:30～10:00

Plenary
Lecture-8

Plenary
Lecture-9

Chairperson: Hiroshi Okada (Japan)

Tending to male factor couples-Gianpiero D. Palermo (USA)
Chairperson: Yoshiharu Morimoto (Japan)

IVM for fertility treatment and preservation of fertility-Seang Lin Tan (Canada)

4. Pre-Congress Workshops:
1) Vitrification
Saturday, September 10 9:00～12:00 (Hana A)
Chairpersons: Stanley Leibo (USA)
Sherman Silber (USA)(1) Lectures
1. Basic science of vitrification

Stanley Leibo (USA)

2. Ovarian reserving (basic research)

Noriko Kagawa (Japan)

3. Ovary transplant: Ovary allotransplantation between

Sherman Silber (USA)(2) Live demonstration
4. The Cryotop® method

Masashige Kuwayama (Japan)

2) Pre-implantation genetic diagnosis (PGD)
Saturday, September 10

14:00～17:00 (Hana A)

Chairpersons: Naoki Aoyama (Japan)
Ilan Tur-Kaspa (USA)(1) Lectures
1. Overview
2. Whole genome amplification is a new basic technology for nextgeneration preimplantation genetic diagnosis on mendelian
inheritance disease
3. To PGD or not to PGD? How many oocytes are needed to PGDwith?
(2) Live demonstration
4. Efficient techniques of PGD for recurrent pregnancy loss (Live)34

Kou Sueoka (Japan)
Suguru Sato (Japan)
Ilan Tur-Kaspa (USA)Naoki Aoyama (Japan)

SCIENTIFIC PROGRAM

5. Concurrent Symposia:
C-1 Management of PCO patients (1)Sunday, September 11 10:30～11:50 (Concord Ballroom A)
Chairperson: Timur Gürgan (Turkey)
Makio Shozu (Japan)
10:30～10:50

C-1. 1Androgenic spectrum concept of PCO morphogenesis-Takahide Mori (Japan)

10:50～11:10

C-1. 2Do phenotypic differences in PCOS affect fertility treatment andoutcomes?
-Rogerio A. Lobo (USA)11:10～11:30

C-1. 3

Laser assisted ICSI for oocytes matured in vitro from PCO patients-Muchsin Jaffar (Indonesia)

11:30～11:50

C-1. 4The use of GnRH antagonist in the controlled ovarian stimulation
for IVF in PCOS patients-Khaled Mahmoud (Tunisia)

C-2 Management of PCO Patients (2)Sunday, September 11 13:20～15:00 (Concord Ballroom A)
Chairpersons: Rogerio A. Lobo (USA)
Khaled Mahmoud (Tunisia)
13:20～13:40

C-2. 1

Polycystic ovary syndrome is associated with metabolic
abnormalities in the purine mononucleotide pathway andtricarboxylic acid cycle-Makio Shozu (Japan)

13:40～14:00

C-2. 2

How to get pregnancy in women with PCOS
-Chii-Ruey Tzeng (Taiwan)

14:00～14:20

C-2. 3ART Management in PCO-Toshiro Kubota (Japan)

14:20～14:40

C-2. 4

PCO and ovarian drilling-Timur Gürgan (Turkey)

14:40～15:00

C-2. 5

PCO and transvaginal hydrolaparoscopic ovarian drilling(THLOD)-Hiroaki Shibahara (Japan)

C-3 Controlled ovarian stimulation (1)Sunday, September 11 15:20～16:40 (Concord Ballroom A)
Chairpersons: Bruno Lunenfeld (Israel)
Milton Leong (Hong Kong)
15:20～15:40

C-3. 1

Available tools to personalize ovarian stimulation - Dose
prediction/protocol selection-Bruno Lunenfeld (Israel)

35

SCIENTIFIC PROGRAM

15:40～16:00

C-3. 2

Advances in ultrasound monitoring of IVF cycles by automatedvolume measurement- Baris Ata (Canada)

16:00～16:20

C-3. 3

Volmetric measurements of the follicular development-Maximilian Murtinger (Austria)

16:20～16:40

C-3. 4

Ovarian stimulation and endometrial receptivity-Pak-Chung Ho (Hong Kong)

C-3 Controlled ovarian stimulation (2)Sunday, September 11 16:40～18:00 (Concord Ballroom A)
Chairpersons: Svend Lindenberg (Denmark)Minoru Irahara (Japan)
16:40～17:00

C-3. 5

Minimal ovarian stimulation (mini-IVF) for IVF utilizing
vitrification and cryopreserved embryo transfer-John Zhang (USA)

17:00～17:20

C-3. 6

Poor responder patients undergoing IVF treatment-Svend Lindenberg (Denmark)17:20～17:40

C-3. 7Prediction and prevention of the ovarian hyperstimulation
syndrome (OHSS) – an evidence-based approach-Hassan Sallam (Egypt)

17:40～18:00

C-3. 8What should we prefer for IVF – the agonist or antagonist?
-Zion Ben-Rafael (Israel)

C-4 Treatment of benign tumor prior to IVF-ET (1)Sunday, September 11 10:30～11:50 (Concord Ballroom B)
Chairpersons: Zion Ben-Rafael (Israel)
Hassan Sallam (Egypt)
10:30～10:50

C-4. 1

Evidence-based management of endometriosis-associatedinfertility-Hassan Sallam (Egypt)

10:50～11:10

C-4. 2

Surgery of endometriosis in infertility: state of the art!
-Issam Lebbi (Tunisia)

11:10～11:30

C-4. 3

11:30～11:50

C-4. 4

Surgical management of recurrent endometrioma: prior to IVF ornot-Hiroshi Nabeshima (Japan)
Management of hydrosalpinges prior to IVF-ET-Victor Gomel (Canada)

C-5 Treatment of uterine myoma or adenomyosis prior to IVF-ET (2)Sunday, September 11 13:20～15:00 (Concord Ballroom B)
Chairpersons: Victor Gomel (Canada)Issam Lebbi (Tunisia)

36

SCIENTIFIC PROGRAM

13:20～13:40

C-5. 1

Surgery of intra-mural leiomyomas in infertile patients-Jean-Bernard Dubuisson (Switzerland)

13:40～14:00

C-5. 2

Myomectomy during pregnancy-Yuji Hiramatsu (Japan)

14:00～14:20

C-5. 3

Uterine fibroids and IVF- what is the controversy?
-Zion Ben-Rafael (Israel)

14:20～14:40

C-5. 4

A novel operative treatment of severe adenomyosis prior toembryo transfer-Hisao Osada (Japan)

14:40～15:00

C-5. 5Different surgical approaches for myomectomy and foradenomyosis-Michael DeRosa (USA)C-6 Oncofertility - Fertility preservation in women with malignant disease (1)Sunday, September 11 15:20～16:40 (Concord Ballroom B)
Chairpersons: Claus Yding Andersen (Denmark)Takeshi Maruo (Japan)
15:20～15:40

C-6. 1The oncofertility: consequences of a previous oncological treatments
and possibilities-Rufino García-Otero Reina (Spain)

15:40～16:00

C-6. 2

Fertility Sparing in Stage Ia2 to Ib1-Cervix Carcinoma-Werner Lichtenegger (Germany)

16:00～16:20

C-6. 3

Abdominal radical trachelectomy in our experience of 126 cases-Takuma Fujii (Japan)

16:20～16:40

C-6. 4Laparoscopic radical trachelectomy-Masaaki Ando (Japan)

C-7 Oncofertility - Fertility preservation in women with malignant disease (2)Sunday, September 11 16:40～18:00 (Concord Ballroom B)
Chairpersons: Klaus Diedrich (Germany)
Kazunori Ochiai (Japan)
16:40～17:00

C-7. 1

Cryopreservation of human oocytes for cancer patients in Japan-Shokichi Teramoto (Japan)

17:00～17:20

C-7. 2Cryopreservation of human ovarian tissue, clinical data and scientificconsiderations-Claus Yding Andersen (Denmark)17:20～17:40

C-7. 3

Consevative therapy for borderline ovarian cancer-Liong Tulusan (Germany)

17:40～18:00

C-7. 4

Infertility, ovulation induction treatments and the incidence of breast,

37

SCIENTIFIC PROGRAM

ovarian and endometrial cancers- thirty years of follow up- Bruno Lunenfeld (Israel)

C-8 ART-cryopreservation: Is oocyte cryopreservation a revolution in IVF?
Sunday, September 11 10:30～11:50 (Concord Ballroom C)
Chairpersons: Stanley Leibo (USA)Masashige Kuwayama (Japan)
10:30～10:50

C-8. 1Highly efficient and safe vitrification using hydroxypropyl
cellulose as a macromolecular supplement for cryopreservation ofoocytes and blastocysts-Luis Arturo Ruvalcaba Castellón (Mexico)

10:50～11:10

C-8. 2

Role and possibility of vitrification in human oocyte-Masashige Kuwayama (Japan)

11:10～11:30

C-8. 3

Blastulation and pregnancy rates after vitrified human zygote
culture for 4 days: Preliminary results-Safaa Al Hasani (Germany)

11:30～11:50

C-8. 4Cryopreservation in human ART-Stanley Leibo (USA)C-9 Children follow-up in ART(1)Sunday, September 11 13:20～14:40 (Concord Ballroom C)
Chairpersons: Fethi Zhioua (Tunisia)
Osamu Ishihara (Japan)
13:20～13:40

C-9. 1

Evaluate the IVF success rates of accumulated pregnancy rates
per oocyte retrieval-Jia-Yin Liu (China)

13:40～14:00

C-9. 2

Genetics and health of children born from cryopreserved oocytes-Ilan Tur-Kaspa (USA)

14:00～14:20

C-9. 3What has happened on epigenetics in ART children?
-Yukiko Katagiri (Japan)

14:20～14:40

C-9. 4Pregnancy and child outcome after assisted reproductive
technologies-Dov Feldberg(Israel)

C-10 Children follow-up in ART(2)Sunday, September 11
15:00～16:20 (Concord Ballroom C)
Chairpersons: Hideaki Kanzaki (Japan)
Dov Feldberg (Israel)
15:00～15:20

C-10. 1

Just Twins? Perinatal data from multiple pregnancies-Peter Brockerhoff (Germany)

15:20～15:40

C-10. 2Perinatal outcomes of ART pregnancies-Fethi Zhioua (Tunisia)

38

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15:40～16:00

C-10. 3

Outcome of IVM babies-Ri-Cheng Chian (Canada)

16:00～16:20

C-10. 4

Perinatal outcome of vitrified human blastocysts in 11 years
experience (5434 attempted cycles) including the rate ofmonozygotic twinning (MZT)-Tetsunori Mukaida (Japan)

C-11 Laboratory advances in IVF (1)Sunday, September 11 16:20～17:40 (Concord Ballroom C)
Chairpersons: Pierre Vanderzwalmen (Belgium)
Gábor Vajta (Australia)
16:20～16:40

C-11. 1Novel embryo culturing and monitoring systems-Artur Bernard (Hungary)

16:40～17:00

C-11. 2

Effects of cumulus cells removal after 6 h co-incubation of gametes
on the outcomes of human IVF-Guoning Huang (China)

17:00～17:20

C-11. 3

An integral view on therapeutic options and lab-techniques forindividualized therapy in IVF-Nicolas Zech (Austria)

17:20～17:40

C-11. 4

How to manage the modern IVF laboratory-Zsolt Peter Nagy (USA)C-12 Laboratory advances in IVF (2)Sunday, September 11 13:20～14:40 (Hana A)
Chairpersons: Yoshihiko Hosoi (Japan)
Zsolt Peter Nagy (USA)
13:20～13:40

C-12. 1

Chemical substances exposure on IVF-ET environment-Tsunehisa Makino (Japan)

13:40～14:00

C-12. 2

Recent insights into the oocyte maturation in ART-Tae-Ki Yoon(Korea)

14:00～14:20

C-12. 3Use of high concentrations of cryoprotectants: is it a justified
argument to prefer slow freezing instead of vitrification?
-Pierre Vanderzwalmen (Belgium)

14:20～14:40

C-12. 4

Embryo culture: Can we perform better than Nature?
-Gábor Vajta (Australia)

C-13 Controlled ovarian stimulation (3)Monday, September 12 10:30～11:50 (Concord Ballroom A)
Chairpersons: Herbert Zech (Austria)
Osamu Tsutsumi (Japan)
10:30～10:50

C-13. 1Superovulation/IUI - An Alternative to IVF-Bill Yee (USA)39

SCIENTIFIC PROGRAM

10:50～11:10

C-13. 2Treatment of patients with high FSH with delayed stimulation
using low dose gonadotrophins-Milton Leong (Hong Kong)

11:10～11:30

C-13. 3Use of hCG in ovarian stimulation-Johan Smitz (Belgium)

11:30～11:50

C-13. 4

Follicle stimulation by transdermal application of FSH-Herbert Zech (Austria)

C-14 Advances in basic research and laboratory techniques in IVM (1)Monday, September 12 13:20～14:20 (Concord Ballroom A)
Chairpersons: Yoshiharu Morimoto (Japan)
Ri-Cheng Chian (Canada)
13:20～13:40

C-14. 1

IVM: learning from IVM ?
-Nelly A. Frydman (France)

13:40～14:00

C-14. 2

Quality evaluation of human & mouse IVM embryo andepigenetic evaluation of human born IVM babies-Hiroaki Yoshida (Japan)

14:00～14:20

C-14. 3

Lessons from the mouse follicle culture and in vitro maturation
models-Johan Smitz (Belgium)

C-14 Advances in basic research and laboratory techniques in IVM (2)Monday, September 12 14:20～15:20 (Concord Ballroom A)
Chairpersons: Jie Qiao (China)
Hiroaki Yoshida (Japan)
14:20～14:40

C-14. 4

Laboratory aspect of natural cycle IVF/IVM treatment-Ri-Cheng Chian (Canada)

14:40～15:00

C-14. 5

Cytoplasmic maturation and mitochondrial activity in human IVM-Yoshiharu Morimoto (Japan)

15:00～15:20

C-14. 6

Correct embryo selection improves embryo implantation rates in
IVM cycles- Hai Ying Chen (Canada)

C-15 Clinical outcomes; toward a more successful IVM
Monday, September 12 15:40～17:20 (Concord Ballroom A)
Chairpersons: Johan Smitz (Belgium)
Aisaku Fukuda (Japan)
15:40～16:00

C-15. 1

IVM application in PCOS patients- Jie Qiao (China)

16:00～16:20

C-15. 2

Oocyte maturation from tiny follicles in human ovarian tissue-Shu Hashimoto (Japan)

40

SCIENTIFIC PROGRAM

16:20～16:40

C-15. 3Natural cycle combined with IVM-Jin-Ho Lim (Korea)

16:40～17:00

C-15. 4The effective approach for IVM using Metformin-Aisaku Fukuda (Japan)

17:00～17:20

C-15.5

Clincal implications of IVM-Svend Lindenberg (Denmark)C-16 Embryo development and competency from activation to implantation
Monday, September 12 10:30～12:10 (Concord Ballroom B)
Chairpersons: Jin-Ho Lim (Korea)
Atsushi Azumaguchi (Japan)
10:30～10:50

C-16. 1Morphological aspect-Nelly A. Frydman (France)

10:50～11:10

C-16. 2

Early embryo development: what's critical-Moncef Benkhalifa (France)

11:10～11:30

C-16. 3

Implantation window-Atsushi Azumaguchi (Japan)

11:30～11:50

C-16. 4

Luteal phase support-Svend Lindenberg (Denmark)11:50～12:10

C-16. 5Predictors of IVF outcome with a particular interest in the amountof gonadotropin administered-Peter Kovacs (Hungary)

C-17 ART-Gamete manipulation
Monday, September 12 13:20～14:20 (Concord Ballroom B)
Chairpersons: Moncef Benkhalifa (France)
Atsushi Tanaka (Japan)
13:20～13:40

C-17. 1Whole ooplasmic replacement in Germinal Vesicle oocytes-Akiko Yabuuchi (Japan)

13:40～14:00

C-17. 2

Dynamic genome function and chromosomes abnormalities-Moncef Benkhalifa (France)

14:00～14:20

C-17. 3

A novel trial of nuclear transfer for repairing an aged oocyte-Atsushi Tanaka (Japan)

C-18 Update on male infertility - IMSI/ ICSI/MD-TESE/PGD
Monday, September 12 14:20～15:20 (Concord Ballroom B)
Chairpersons: Sherman Silber (USA)
Hiroshi Okada (Japan)
14:20～14:40

C-18. 1

Male infertility-Sherman Silber (USA)41

SCIENTIFIC PROGRAM

14:40～15:00

C-18. 2

15:00～15:20

C-18. 3

Microdissection Testicular Sperm Extraction (MD-TESE)-Atsumi Yoshida (Japan)
IMSI, already 8 years in ART practice. Where do we stand?
-Pierre Vanderzwalmen (Belgium)

C-19 AMH: Is it essential for assessing ovarian reserve
Monday, September 12 10:30～11:50 (Concord Ballroom C)
Chairpersons: Bunpei Ishizuka (Japan)
Hsin-Fu Chen (Taiwan)
10:30～10:50
C-19. 1
Clinical application of AMH in the reproductive medicine-Yoshimasa Asada (Japan)
10:50～11:10

C-19. 2Chronological aging vs biological aging: AMH as an earlymarker of biological aging-Budi Wiweko (Indonesia)

11:10～11:30

C-19. 3

POF and AMH-Bunpei Ishizuka (Japan)

11:30～11:50

C-19. 4

Dynamic serum AMH levels and AMH-related gene expression
in human ovarian follicles-Hsin-Fu Chen (Taiwan)

C-20 The role of ART in international development (1)Monday, September 12 13:20～14:40 (Concord Ballroom C)
Chairpersons: Willem Ombelet (Belgium)
Sheryl Vanderpoel (Switzerland)
13:20～13:40

C-20. 1

Implementation of the fertility assistance program after the
Wenchuan earthquake in China-Shangwei Li (China)

13:40～14:00

C-20. 2

Cross-border cryo-shipping of vitrified embryos (CCVE):
the newest ART success story-Gautam Allahbadia(India)

14:00～14:20

C-20. 3

Cross-border reproductive care: exploitation or opportunity inthe global quest for a baby?
-Daniel S. Seidman (Israel)

14:20～14:40

C-20. 4

Oocyte donors: prevalence of genetic, infectious diseases andothers-Oriol Coll (Spain)

C-20 The role of ART in international development (2)Monday, September 12

14:40～15:40 (Concord Ballroom C)

Chairpersons: Daniel Seidman (Israel)

Tetsunori Mukaida (Japan)
14:40～15:00

C-20. 5

Vouchers for health: A demand side output-based aid approach
toreproductive health services. A model to pay for infertility
services?

42

SCIENTIFIC PROGRAM-Claus Peter Janisch (Germany)
15:00～15:20

C-20. 6

Global challenges and perspectives-Sheryl Vanderpoel (Switzerland)

15:20～15:40

C-20. 7

Affordable IVF: low cost IVF – The Arusha project-Willem Ombelet (Belgium)

C-21 Repeated implantation failure: how to improve implantation in ART
Tuesday, September 13 10:30～11:50 (Concord Ballroom A)
Chairpersons: Kouichi Takakuwa (Japan)
Yael Gonen (Israel)
10:30～10:50

C-21. 1

How to improve implantation in ART: novel approaches-Yael Gonen (Israel)

10:50～11:10

C-21. 2

Studies on the efficacy of immunotherapy using paternalmononuclear cells for patients with infertility-Kouichi Takakuwa (Japan)

11:10～11:30

C-21. 3

Improvement of implantation rates using autologous peripheral
blood mononuclear cells-Hiroshi Fujiwara(Japan)

11:30～11:50

C-21.4

Prevention of infertility in patients for the antiphospholipid
syndrome and deficiencies of natural anticoagulants-Moritoshi Seki (Japan)

C-22 Pre-implantation genetic screening: an update
Tuesday, September 13 10:30～11:50 (Concord Ballroom B)
Chairpersons: Hidekazu Saito (Japan)
Jiaen Liu (China)
10:30～10:50

C-22. 1PGD for recurrent pregnancy loss
-Atsushi Tanaka (Japan)

10:50～11:10

C-22. 2

Polar body analysis in clinical practice as compared to pre-embryo
morphology for selection of the best for transfer-Wilfried Feichtinger (Austria)

11:10～11:30

C-22. 3

PGD outcome and alternatives for single gene disorder-Kou Sueoka (Japan)

11:30～11:50

C-22.4

Preimplantation genetic diagnosis for aneuploidy using wholegenome amplification and microarray comparative genomic
hybridization-Alan Handyside (UK)C-23 Stem cell and fertility preservation: Where are we at with stem cells?
Tuesday, September 13 10:30～12:10 (Concord Ballroom C)
Chairpersons: Kwang Yul Cha (Korea)
Oktay Kadayifci (Turkey)

43

SCIENTIFIC PROGRAM

10:30～10:50

C-23. 1

Human ES cell and iPS cell derivation: Clinical applications andbiological characterization-Hidenori Akutsu (Japan)

10:50～11:10

C-23. 2Pluripotent stem cell as a source of germ cells - Is it still a fiction?
-Anis Feki (Switzerland)

11:10～11:30

C-23. 3

11:30～11:50

C-23. 4Development of pluripotent stem cells to female germ cells-Hsin-Fu Chen (Taiwan)
Testicular stem cells as sources of fertility preservation and celltherapy-Kwang Yul Cha (Korea)

11:50～12:10

C-23. 5In vitro production of functional sperm in cultured neonatal mousetestes-Takehiko Ogawa (Japan)

6. STGO Session (Tunisian Society of Gynecology and Obstetrics)(Coordinators: Khaled Mahmoud, Fethi Zhioua, Issam Lebbi)
Challenging issues in ART (1)Monday, September 12 16:00～17:00 (Concord Ballroom C)
Chairpersons: Hedi Khairi
Anis Feki
Said Lazrak
16:00～16:20

STGO (1) 1

How to manage hyper-response to COS in IVF-Youssef Boutaleb (Morocco)

16:20～16:40

STGO (1) 2

How to manage poor-responders?
-René Frydman (France)

16:40～17:00

STGO (1) 3

The thin endometrium in ART: What to do?
-Amina Oumziane (Algeria)

Challenging issues in ART (2)Monday, September 12 17:00～18:00 (Concord Ballroom C)
Chairpersons: Mahmoud Kharouf
Moise Fiadjoe
Nabil Ben Zineb
17:00～17:20

STGO (2) 1

How to treat the female causes of repeated implantation failure?
-Khaled Terras (Tunisia)

17:20～17:40

STGO (2) 2

How to manage sperm investigation after ICSI Failure?
-Moncef Benkhalifa (France)

17:40～18:00

STGO (2) 3

How to improve laboratory procedures to enhance implantation?
-Ghaya Merdassi (Tunisia)

44

SCIENTIFIC PROGRAM

7. ISF Session (Israeli Society of Fertility) (Coordinator: Daniel Seidman)
Monday, September 12 15:40～17:40 (Concord Ballroom B)
Chairpersons: Arye Hurwitz
Martha Dirnfeld
15:40～16:00

ISF 1.

Fertility enhancement in patients with adenomyosis using MRI
guided HIFUS-Yaron Rabinovici (Israel)
Gonadotropin-releasing hormone agonist trigger: the way toeliminate OHSS
-Arye Hurwitz (Israel)

16:00～16:20

ISF 2.

16:20～16:40

ISF 3.

From endometrial injury to IMSI: New approaches for repeated IVF
failure-Martha Dirnfeld (Israel)

16:40～17:00

ISF 4.

High ART success rates when financial constraints are completely
lifted-Daniel Seidman (Israel)

17:00～17:20

ISF 5.

Autologous transplantation of very thin ovarian fragments which
preserve the ovary's main cortex structure lead to successful
pregnancy-Ariel Revel (Israel)

17:20～17:40

ISF 6.

How should we consult candidates for fertility preservation due to amalignant disease regarding their expected ovarian response to
COH?
-Shevach Friedler (Israel)

8. APART Session: MSCs - From basic research to clinical applications
Monday, September 12 13:20～15:00 (Hana A)
Chairpersons: Yuji Takehara (Japan)
Byung-Rok Do (Korea)
13:20～13:40

APART 1.

Clinical efficacy of adipose MSCs in human-Byung-Rok Do (Korea)

13:40～14:00

APART 2.

Human lipoaspiration-Ken Nakama (Japan)

14:00～14:20

APART 3.

Vitrification of adipose MSCs
-Noriko Kagawa (Japan)

14:20～14:40

APART 4.

Preparation of adipose-derived stem cells-Rie Yamadera (Japan)

14:40～15:00

APART 5.

Discussion about clinical applications of adipose MSCs
-Yuji Takehara (Japan)

45

SCIENTIFIC PROGRAM

9. Oral Communications
Sunday, September 11

10:30～12:00(Hana A)

Chairpersons:
O-001

Daniel Bodri (Hungary)
Atsumi Yoshida (Japan)
Does male reproductive tract CD52 (mrt-CD52) prevent complement activation via
binding C1q?
○L. Hardiyanto, A. Hasegawa, S. Komori
Hyogo College of Medicine, Kobe, Japan

O-002

Technology Research of Prostatic Urethral Irrigation and Drainage Catheter
Injection○Weidong Huang
Xinjiang Jiayin Hospital Center for Reproductive Medicine, Urumqi, China

O-003

Measurement of reactive oxygen species in neat and washed sperm, is there any
difference?
○Mohammadreza Moein, Nasim Tabibnejad, Jalal Ghasemzadeh
Yazd Research and Clinical Center for Infertility, IranChairpersons:O-004

Naoki Takeshita (Japan)
Naoki Aoyama (Japan)
Whole Genome Amplification is efficient for CTG repeats length detection of PGD for
DM1
1,2

2

2

2

2

2

○Tomoyoshi Sakurai , Kou Sueoka , Kaori Takahashi , Suguru Sato , Kenji Sato ,Yasunori Yoshimura

O-005

1

Saitama municipal Hospital Department of Obstetrics and Gynecology, Department of Obstetrics and Gynecology, Saitama, Japan,

2

Keio University School of Medicine, Tokyo, Japan

Reducing multiple pregnancies by selection of a single chromosomally normal
blastocyst for transfer using array CGH (aCGH) analysis of 24 chromosomes within
24 hours
1

2

1

2

1

○Zhihong Yang , Xiaohong Liu , Shala Salem , Jiaen Liu , Rifaat Salem
1

O-006

Pacific Reproductive Center, Torrance, California USA, 2Beijing Jiaen Hospital, 29 Zhichun lu, Beijing 10083, China

Preimplantation genetic screening of eggs and blastocysts with CGH-microarray:clinical experience with the first 51 cases
1

1

1

1

1

○Stuart Lavery , Ben Lavender , Paul Knaggs , Anastasia Mania , Geoffrey Trew , Dagan
2
Wells
1

IVF Hammersmith, Hammersmith Hospital, London UK, 2Reprogenetics Oxford UK

Sunday, September 11

15:00～18:00(Hana A)

Chairpersons:
O-007

Budi Wiweko (Indonesia)
Shu Hashimoto (Japan)
Most Motile Sperm Can Be Separated at the Bottom of the Microfluidic Channel of a
Plastic Microfluidic Sperm Sorter
1○Koji Matsuura , Keiji Naruse

2

1

Research Core for Interdisciplinary Sciences, Okayama University, Okayama, Japan, 2Cardiovascular Physiology, Graduate
School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan

O-008

Polymerization of Insulin-Like Growth Factor-Binding Protein-1 (IGFBP-1)
Potentiates IGF-I Actions in Placenta

46

SCIENTIFIC PROGRAM
1

2

2

2

2

○M Kabir-Salmani , H Shibuya , K Sakai , Y Wachi , M Iwashita
1

2

National Institute of Genetic Engineering and Biotechnology,Tehran, Iran, Department of Obstetrics and Gynecology, Kyorin
University School of Medicine, Tokyo, Japan

O-009

MicroRNAs associated with human embryo implantation defects
1

2

3

4

○Ariel Revel , Hanna Achache , Juliet Stevens , Smith Yoav
1

2

Hadassah University hospital. Jerusalem, Israel, Institute for Drug Research, School of Pharmacy, Faculty of medicine, The
Hebrew University of Jerusalem, Jerusalem, Israel, 3University of Oxford, Oxford, United Kingdom, 4Genomic Data Analysis
Unit, The Hebrew University of Jerusalem, Jerusalem, Israel

O-010

Pre-ovulation leptin serum as a marker for endometrial receptivity
11○Andon Hestiantoro , Marly Susanti , Atikah Barasila

2

1Division Reproductive Immuno-endocrinology, Department of Obstetrics and Gynecology, Faculty of Medicine University ofIndonesia, Dr. Cipto Mangunkusumo., 2Departmen of Hystology, Faculty of Medicine, University of Indonesia, Jakarta,
Indonesia

Chairpersons:O-011

Daniel Bodri (Hungary)
Akira Kuwahara (Japan)
Application of Mechanical Stimuli Using a Microfluidic Air Actuation System:
Dynamic Embryo Culture
1

1○Koji Matsuura , Yuka Kuroda , Keiji Naruse

2

1

Research Core for Interdisciplinary Sciences, Okayama University, Okayama, Japan, 2Cardiovascular Physiology, Graduate
School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan

O-012

Caffeine treatment influences in vivo and in vitro fertilization, embryo development
and morphology in mice
1

1

1

1

○Yoshimasa Yokota , Mikako Yokota , Hidemi Yokota , Setsuko Sato , Yasuhisa Araki
1

2

2

Yokota Maternity Hospital, The Institute for Advanced Reproductive Medical Technology, Gunma, Japan

O-013

Dynamic analysis of compaction initiation in human embryos using time-lapse
cinematography○Kyoko Iwata, Keitaro Yumoto, Akifumi Imajo, Yumiko Iba, Yasuyuki Mio
Mio Fertility Clinic, Yonago, Japan

O-014

Chromosomal analysis of cleavage embryos and blastocysts derived fromtripronuclear zygotes after ICSI.
○Shimpei Mizuta, Nobuhiko Kataoka, Hiromi Hashimoto, Yasushi Kuroda, Shoji Kokeguchi,
Masahide Shiotani
Hanabusa Women's Clinic, Hyogo, Japan

Chairpersons:O-015

Muchsin Jaffar (Indonesia)
Fumihito Aono (Japan)
THE DYNAMIC PROCESS OF SPERM PENETRATION OF THE HUMAN OOCYTEANALYZED USING TIME-LAPSE CINEMATOGRAPHY○Yoshiteru Kai, Kyoko Iwata, Keitaro Yumoto, Akifumi Imajo, Yumiko Iba, Yasuyuki Mio
Mio Fertility Clinic, Yonago, Japan

O-016

Depletion of Plk1 delays cell cycle progression in endometrial cells derived fromendometriosis
1,2

3

4

2

2

Li Tang , Tian-Hua Zhou , Jian-Zhong Sheng , Yan-Ting Wu , Ming-Yue Dong , ○He-Feng
2
Huang
1

The First People's Hospital of Yunnan Province, 2Women's Hospital, Zhejiang University, School of Medicine,
Zhejiang University, School of Medicine, 4University of Calgary, Canada

347

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O-017

Male fetal DNA detection in maternal serum from pregnant cynomolgus monkeys(Macaca fascicularis) in an established breeding colony
1

2

3

3

○Lubna YASMIN , Jun-ichiro TAKANO , Yasushi NAGAI , Junko OTSUKI , Tadashi
1
SANKAI
1

Tsukuba Primate Research Center, National Institute of Biomedical Innovation , 2Department of Research Resource
Developments, The Corporation for Production and Research of Laboratory Primates, 3Nagai Clinic

O-018

Clinical outcome of 852 tubal factor infertility patients without ART after
Falloposcopic tuboplasty○Shoji Kokeguchi, Nobuhiko Kataoka, Seiji Ogata, Satoshi Yamada, Yukiko Matsumoto,
Masahide Shiotani
Hanabusa Women's Clinic, Hyogo, Japan

Monday, September 12

10:30～12:15(Hana A)

Chairpersons:O-019

Saffa Al Hassani (Germany)
Tesunori Mukaida (Japan)
Outcomes of vitrified-warmed embryo transfer (FET) cycles: single blastocyst
transfer versus double blastocyst transfer○Maria Milyutina, Elena S. Miadova
Perinatal Medical Center, Moscow, Russia

O-020

Are Mild ART-derived blastocysts more favorable than conventional COH-derived
ones?
1

1

1

1

2

○Yasushi Takai , Ken Ohara , Shigetaka Matsunaga , Masahiro Saito , Osamu Ishihara ,1
Hiroyuki Seki
1

Saitama Medical Center/Saitama Medical University, Saitama, Japan, 2Saitama Medical University, Saitama, Japan

O-021

Progress towards a universal warming method after vitrification
1

1,2

1

1

○Aniko Reichart , Gabriella Uhereczky , Miklos Sipos , Vince Forgacs , Gabor Vajta
122

Forgacs Institute, Budapest, Hungary, BGI Shenzhen, Yatian District, Shenzhen, China

Chairpersons:O-022

Nicolas Zech (Austria)
Masao Jinno (Japan)
Failure of pronucleus formation in in vitro matured and fertilized oocytes from
SOD1-deficient mice
1

1

1

2

○Naoko Kimura , Yasuko Sato , Manami Suenaga , Satoshi Tsunoda , Junichi Fujii

2

1

Laboratory of Animal Reproduction, Graduate School of Agricultural Sciences, Yamagata University, Tsuruoka, Japan,
Department of Biochemistry and Molecular Biology, Graduate School of Medical Science, Yamagata University, Yamagata,
Japan
2

O-023

A randomized double-blind comparative study between two different laser assisted
hatching in the frozen-thawed embryo transfer at the cleavage stage○Lei Ao
Yunnan First People Hospital, Kunming, China

O-024

A trial for observation of chromosome dynamics in human embryos using a live-cell
imaging system
1

1

1

1

1

○Yoshiharu Nakaoka , Shu Hashimoto , Ami Amo , Keijiro Ito , Yoshiharu Morimoto ,2
Kazuo Yamagata
1

IVF Namba Clinic, Osaka, Japan, 2Research Institute for Microbial Diseases, Osaka University

48

SCIENTIFIC PROGRAM

O-025

Autologous transplantation of very thin ovarian fragments which preserve the ovary's
main cortex structure lead to successful pregnancy
1

1

1

1

1

○Ariel Revel , Meital Lebovich , Alex Simon , Neri Laufer , Einat Eizenmann , Eduardo
2
Mitrani1

Hadassah University hospital. Jerusalem, Israel, 2The Alexander Silberman Institute of Life Sciences, hebrew university,
Jerusalem, Israel

Sunday, September 12

15:20～18:05(Hana A)

Chairpersons:O-026

Yealin Frank (USA)Yumi Nagata (Japan)
Important implications of advanced glycation end-products (AGE) in poor ART
outcomes and a novel successful therapy for very severe ART patients withsitagliptin possibly by decreasing AGE.
1

2

1

1

○Masao Jinno , Masayoshi Takeuchi , Aiko Watanabe , Jun Hirohama ,11
Naohisa Hatakeyama , Rie Hiura
1

Women's Clinic Jinno, Tokyo, Japan, 2Kanazawa Medical College, Ishikawa, Japan

O-027

A novel long protocol of GnRH agonist and hMG regimen: a dramatical increase inpregnancy rate by induction of diminished but significant mid-cycle LH surge.
○Masao Jinno, Aiko Watanabe, Jun Hirohama, Naohisa Hatakeyama, Rie Hiura,
Rika Nishiyama
Women's Clinic Jinno

O-028

GnRHa: a promising alternative of HCG to trigger ovum maturation in minimal
stimulation cycle
Ze Wu, Bo Deng, Yonggang Li, Yunxiu Li, ○Yanping li
Department of Reproduction and Genetics; Reproductive Medicine Centre, The First Peoples Hospital of Yunnan Province, China

O-029

The observation of effect of two different COH protocols on the IVF-ET outcome ofaged patientsBo Deng, Ze Wu, Yanping Ma, ○Yonggang Li
Department of Reproduction and Genetics; Reproductive Medicine Centre, The First Peoples Hospital of Yunnan Province, China

Chairpersons:O-030

Maximlian Murtinger (Austria)
Moritoshi Seki (Japan)
EFFICACY OF SEQUENTIAL TREATMENT PROTOCOL WITH HIGHLY PURIFIEDURINARY FSH AND RECOMBINANT FSH FOR CONTROLLED OVARIANSTIMULATION○Hong Ye, Guoning Huang, Li Pei, Pinghong Zeng, Xiu Luo
1

Chongqing Genetic and Reproductive Institute, Chongqing Obstetrics and Gynecology Hospital, China

O-031

Application of elective single embryo transfer(e-SET)○Yonggang Li, Yanping Ma, Ze Wu, Lian Deng, Bo Deng, Mengying Gao
Dep of Reproduction and Genetics, Yunnan first people hospital, Kunming, China

O-032

A Novel function of Human Pinopodes by Expressing L-Selectin Ligand during
Window of Implantation
1

2

3

4

○M Kabir-Salmani , R Nejatbakhsh , H Hosseini , E Dimitriadis , M Iwashita
125

National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran., Dept. Anatomical Sciences, Shaheed
Beheshti Medical University, Tehran, Iran, 3Molecular and Cellular Biology Research Center, Shaheed Beheshti Medical
University, Tehran, Iran, 4Prince Henry's Institute of Medical Research, Australia., 5Dept. Ob/Gyn, Kyorin Medical University,
Tokyo, Japan

49

SCIENTIFIC PROGRAM

O-033

Obstetric outcomes of ART pregnancies in women over 35 years old○Xiaohui Tang, Tomoko Adachi, Setsuko Nakayama, Yoshiharu Takeda, Hideki Sakamoto,
Masao Nakabayashi
Department of Obstetrics and Gynecology, Aiiku Maternal and Child Health Center, Aiiku Hospital, Tokyo, Japan

Chairpersons:O-034

Yan Ping Kuwan (China)
Yasuyuki Mio (Japan)
The age of 35 years is the critical age for successful TESE-ICSI in nonobstructive
azoospermic patients with normal karyotype and nonmosaic Klinefelter syndrome.
1

1

2,3

2,3

○Hiroshi Okada , Yoshitomo Kobori , Mitsunobu Koshida , Ken-Ichi Tatsumi ,3,45Kazutaka Terai , Osamu Maruyama
1

Department of Urology, Dokkyo Medical University Koshigaya Hospital, Saitama, Japan, 2Koshida Clinic, 3Umegaoka
Women's Clinic, 4Department of Urology, Shakaihoken Kamata General Hospital, 5Department of Urology, Juntendo
University Faculty of Medicine

O-035

Comparing study of effectiveness of nifedipine and magnesium sulfate for acutetocolysis of preterm labor and threatened preterm labor
1

2

3

4

5

○Fariba Nanbakhsh , Farzaneh Broomand , Zahra Yekta , Rita Doosti , Pooya Mazloomi
1

Urmia University of Medical Sciences,Urmia,IRAN, 2Urmia University of Medical Sciences,Urmia,IRAN, 3Urmia Universityof Medical Sciences,Urmia,IRAN, 4Urmia University of Medical Sciences,Urmia,IRAN, 5Urmia University of Medical
Sciences, Urmia, IRAN

O-036

The Change of Serum Anti-Mullerian Hormone (AMH) Level by Chemotherapy andOperation in Premenopausal Women with Gynecological Cancer○Masaru Hayashi, Akiko Shoda, Nobuaki Kosaka, Yoshiko Moshiduki, Ichio Fuykasawa
Department of Obstetrics and Gynecology, Dokkyo Medical University, Tochigi, Japan

Sunday, September 13

10:30～11:45(Hana A)

Chairpersons:O-037

Mette Munk (Denmark)Shoji Kokeguchi (Japan)
Alternative treatment options for patients with ovarian insufficiency based on Natural
Cycle IVF
○Markus Nitzschke
Milagro Kinderwunschzentrum Bodensee AG, Kreuzlingen, Switzerland

O-038

CORRELATION OF BODY MASS INDEX WITH OUTCOME OF IN VITROFERTILIZATION IN A DEVELOPING COUNTRY○Neeta Singh, Prerna Gupta, Suneeta Mittal, Neena Malhotra, Anupama Bahadur
All India Institute of Medical Sciences, New Delhi, India

O-039

Bilateral ovarian endometriomas removal does not affect the IVF outcome○Andrej Vogler, Martina Ribic Pucelj, Irma Virant Klun
Department of Obstetrics and Gynaecology, University Medical Centre, Ljubljana, Slovenia

O-040

The new stripping technique for low protrusion rate uterine myoma in TCR○Toshimichi Oki, Chie Oki, Toshihiko Kawamura, Akiko Gibo, Hideki Yamasaki,
Tsutomu Douchi
Wemen's Medical Center, Kagoshima Univesity Hospital, Kagoshima, Japan

O-041

Evaluation of short-term and long-term outcome of tubal conservation in thetreatment of tubal ectopic pregnancy○Toshimichi Oki, Yukiko Nakajou, Chie Oki, Akiko Gibo, Toshihiko Kawamura, Tsutomu
Douchi
Wemen's Medical Center, Kagoshima Univesity Hospital, Kagoshima, Japan

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SCIENTIFIC PROGRAM

O-042

2000IU hCG in high responders does not affect the outcomes of IVF
1

1

1○YanPing Kuang , Yun Wang , QiFeng Lyv , John Zhang

2

Department of Assisted Reproductive, Shanghai Ninth People‘s Hospital Affiliated Shanghai JiaoTong University School of
Medicine, Shanghai, China; 2 New Hope Fertility Center, New York, U.S.
1

10. Poster Presentaions
Poster viewing time (Hana B C D)
Sunday, September 11 10:00～10:30, 13:00～13:20
Monday, September 12 10:00～10:30, 13:00～13:20
P-001

The influence of crowding on Leydig cell, weight gain, testosterone and cortisolelevels in mice
○Maryam Ghasemi, Farzad Rajaei
Department of Anatomy, Qazvin University of Medical Sciences, Qazvin, Iran

P-002

Identification of basic reprogramming factors associated with a pluripotent potential
of in vitro cultured spermatic stem cells in domestic animals.
1

1,2

3

1

1

Sung-Min Kim , Mayako Fujihara , Sadeep Goel , Mahesh Sahare , Naojiro Minami ,1
1
Masayasu Yamada , ○Hiroshi Imai
1

Kyoto University, Kyoto, Japan, 2Smithsonian Coservation Biology Institute, Front Royal, Virginia, USA,
Center for Cellular and Molecular Biology, Hyderabad, India

3

P-003

Effect of diet contain sesame seed on the rat testis
121Javad Amini mohabadi , ○Hassan Hassani Bafrani , Morad Pasha Eskandari Nasab ,1
3
4
Mohammad Hossein Shahir , Hossein Nikzad , Aliakbar Taherian
1

Zanjan University, 2Anatomical Research Center, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran,
Department of Anatomy & Embryology, Scientific Director of the IVF Lab, School of Medicine, Kashan University of
Medical Sciences, Kashan, Iran, 4Department of Anatomy & Embryology, Scientific Director of the IVF Lab, School of
Medicine, Kashan University of Medical Sciences, Kashan, Iran
3

P-004

Analysis of Y chromosome microdeletions in Kashanian infertile males
1

2

1

3

○Mahnaz Torfeh , Hassan Hassani Bafrani , Ebrahim Sakhinia , Mahdi Rohani
1Department of Biochemistry and genetics, Faculty of Medicine, Tabriz University of Medical Sciences, 2Anatomical ResearchCenter, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran. , 3Department of Bacteriology, PastureInstitute of Iran

P-005

The usefulness of testicular sperm extraction with the Trucut biopsy needle○Syuichi Iida, Masakuni Suzuki, Ikuo Tachibana, Sigetomo Takahashi, Takahiro Noda,
Osamu Fuzii
M, Suzuki's Memorial Hospital, Miyagi, Japan

P-006

An evaluation of the confounding effect of sperm abnormalities on pregnancy andimplantation rates and miscarriage rates following in vitro fertilization- embryotransfer using sibling oocytes○Jung K Choe, Jerome H Check, Theresa Jamison
UMDNJ, Robert Wood Johnson Med. School at Camden, Cooper Hosp./Univ. Med. Cntr., Dept. OB/GYN, Div. Repro. Endo.
& Infertility, Camden, NJ, U.S.

P-007

Testicular sperm retrieval using a new multiple-holes puncture needle○Hong-Hua Wang, Li-Yi Cai, Hong-Ying Yu, Jing-Ying Xiang, Lin-Qing Hu, Xiao-Jin Zhou
Department of Reproductive Medicine, the Affiliated Wuxi Hospital for Maternal & Childers Health Care of Nanjing Medical
University, China

51

SCIENTIFIC PROGRAM

P-008

The impact of body mass index on sperm recovery and serum reproductive hormone
levels in an infertility setting -An analysis of 445 azoospermic cases
1

1

2○Hatsuki Hibi , Tadashi Ohori , Yoshiaki Yamada , Yoshimasa Asada

3

1

Kyoritsu General Hospital, Nagoya, Japan, 2Department of Urology, Aichi Medical University School of Medicine, 3Asada
Lady's Clinic, Aichi, Japan

P-009

The frequency of males with sperm with low hypoosmotic swelling test scores (which
prevents morphologically normal embryos from implanting) in couples having in vitro
fertilization-embryo transfer○Gabrielle Citrino, Jerome H Check, Jung K Choe, Ann DiAntonio
UMDNJ, Robert Wood Johnson Med. School at Camden, Cooper Hosp./Univ. Med. Cntr., Dept. OB/GYN, Div. Repro. Endo.
& Infertility, Camden, NJ, U.S.

P-010

Effects of several culture conditions on the primordial germ cell proliferation
1

2○Zohreh Makoolati , Mansoureh Movahedin , Mehdi Forouzandeh-Moghadam

3

1

Department of Anatomical sciences, Medical Sciences Faculty, Fasa University, Fasa, , 2Department of Anatomical sciences,
Medical Sciences Faculty, Tarbiat Modares University, Tehran, Iran , 3Department of Biotechnology, Medical Sciences
Faculty, Tarbiat Modares University, Tehran, Iran

P-011

Early development of in vitro aged eggs after intracytoplasmic sperm injection
1

1

1

1

○Gaku Shimoi , Masato Hayashi , Yuichi Kameyama , Ryoichi Hashizume ,2
1
Ken-ichi Kudoh , Masao Ito
1

Faculty of Bioindustry, Tokyo University of Agriculture, Hokkaido, Japan, 2School of Veterinary Medicine, Kitasato
University, Aomori, Japan

P-012

One step closer to the development of a rapid bioassay to determine if adequate
luteal phase progesterone supplementation is provided during in vitro
fertilization-embryo transfer cycles
1

1

2

1

○Ann DiAntonio , Jerome H Check , Maya D Srivastava , Rachael Cohen , Ebony Dix

1

1

1UMDNJ, Robert Wood Johnson Med. School at Camden, Cooper Hosp./Univ. Med. Cntr., Dept. OB/GYN, Div. Repro.
Endo. & Infertility, Camden, NJ, 2Department of Medicine, Division of Allergy and Immunology, SUNY at Buffalo, Buffalo,
NY, U.S.

P-013

Effect of ovarian induction on the ultrastructure of corpus luteum during luteal phaseat implantation period
1

2

3

4

○Mandana Beigi Boroujeni , Nasim Beigi Boroujeni , Mojdeh Salehnia , Elahe Marandi ,4
5
Sadegh Rezapour , Masoud Beigi Boroujeni
1

Department of Anatomy, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran, 2Department ofclinical science, School of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran., 3Department of
Anatomy, School of Medicine, Tarbiat Modares University, Tehran, Iran, 4School of Medicine, Lorestan University of Medical
Sciences, Khorramabad, Iran, 5Department of Biology, School of Basic Science, Shahrekord University, Shahrekord, Iran

P-014

Autophagy in the ovarian granulosa cells○Eri Ishida, Hirohiko Tani, Miwa Shimizu, Akiyoshi Urano, Takakazu Saito, Hidekazu Saito
National Center for Child Health and Development, Department of Women's Health, Division of Reproductive Medicine andInfertility, Tokyo, Japan

P-015

The promoter -1031(T/C) polymorphism in tumor necrosis factor-α associated withpolycystic ovary syndrome1

12○Kwang-Hyun Baek , Ji-Hyun Yun , Jin-Woo Choi , Hyo Young Jeoung
1

3

Department of Biomedical Science, CHA Stem Cell Institute, CHA University, CHA General Hospital, Seoul, Korea, 2St.
Paul's School, Concord, NH, USA, 3Department of Obstetrics and Gynecology, CL Hospital, Gwangju, Korea

52

SCIENTIFIC PROGRAM

P-016

Association between INS-VNTR polymorphism and polycystic ovary syndrome in a
Korean population
1

1

2

2

○Bum-Chae Choi , Sang Jin Song , Ji-Hyun Yun , Bon-Hee Gu , Kwang-Hyun Baek
1

2

2

Department of Obstetrics and Gynecology, CL Hospital, Gwangju, Korea, Department of Biomedical Science, CHA Stem
Cell Institute, CHA University, CHA General Hospital, Seoul, Korea

P-017

Application of non-elective blastocyst culture
Mengying Gao, ○Yanping Ma, Yonggang Li, Ze Wu , Lian Deng, Bo Deng
The Department of Reproduction and Genetics, Yunnan first people hospital, Kunming, P.R. China

P-018

Oocyte activation with Ca ionophore A23187 and roscovitine for ovulated mouse
oocytes to produce haploid parthenogenones○Yuya Yano, Yuri Yamamoto, Yu Tanaka, Kenji Hinokio, Akira Kuwahara, Minoru Irahara
Department of Obstetrics and Gynecology, Institute of Health Biosciences, University of Tokushima Graduate School,
Tokushima, Japan

P-019

The Availability of Recombinant FSH (recFSH) in Minimum Ovarian Stimulation by
the Deference of the Anti-Müllerian Hormone (AMH) Level
1

1

1

1

2

○Yuki Nagase , Miki Ikegami , Maiko Yoshioka , Tomijirou Nishihara , Mitsuru Usui ,1Toshiki Matsuura
1

Kaba Clinic Reproduction Center, Hamamatsu, Japan, 2Kyoritsu Juzen Hospital, Hamamatsu, Japan

P-020

Results of 1027 office-based diagnostic hysteroscopies before IVF and evaluation
the pregnancy and take home baby percentages between patients with normal andabnormal uterine findings.
Selcen Bahadir, ○Mujdegul. Z Karaca, Sertac Batioglu
Zekai Tahir Burak Women's Health Education and Research Hospital, Ankara, Turkey.

P-021

Serum levels of macrophage inhibitory cytokine 1 (MIC 1) as a predictor of
miscarriage in early pregnancy of women treated for repeated reproductive failures○Daniela. N Baltadzhieva, Kalinka. L Penkova, Pepa. A Angelova, Meglena. M Metodieva
Center for Reproductive Health "Nadejda", Sofia, Bulgaria

P-022

A drop in serum estradiol the day after human chorionic gonadotropin (hCG) shot
does not adversely affect pregnancy rates per embryo transfer in women with very
decreased oocyte reserve○Ann DiAntonio, Jung K Choe, Jerome H Check
UMDNJ, Robert Wood Johnson Med. School at Camden, Cooper Hosp./Univ. Med. Cntr., Dept. OB/GYN, Div. Repro. Endo.
& Infertility, Camden, New Jersey, U.S.

P-023

Analysis of the limitations for the numbers of attempts of infertility treatment using
ART.
○Hiroaki Shibahara, Tatsuya Suzuki, Kenro Chikazawa, Tomoe Ikeda, Yuki Hirano,
Mitsuaki Suzuki
Department of Obstetrics and Gynecology, School of Medicine, Jichi Medical University, Tochigi, Japan

P-024

The predictive value of AMH for ovary reserve and response during IVF-ET○Xiaomei Zhang
The reproductive medicine center of Subei People's hospital, China

P-025

Whether Day 3 FSH/LH ratios is a good predictor of IVF prognosis in GnRH
antagonist protocol?
1

2

1

1

1

○K.H. Lee , J.D. Cho , H.G. Sun , S.K. Kim , J.H. Lee , Ilhae Park1

2

1

Mamapapa&baby OB&GY, Ellemedi Infertility Clinic, Changwon, Gyoung Nam, repubilic of Korea

53

SCIENTIFIC PROGRAM

P-026

Luteal blood flow in patients undergoing GnRH agonist long protocol○Kumiko Mizumoto, Akihisa Takasaki, Maki Okada, Yuuko Sakaguchi,
Katsunori Shimamura, Hitoshi Morioka
Saiseikai Shimonoseki Hospital, Yamaguchi, Japan

P-027

Comparison of the effect of GnRHa long and short protocol in older women during
IVF/ICSI○Xun Zeng, Shangwei Li, Lang Qing, Xiaohong Li, Han Hu
Reproductive Medical Center of West China 2nd Hospital,Sichuan,China

P-028

Small amount of testosterone administration improves ovarian response to ovulationinduction in poor responders
1

1

1

1

2

○Kohzo Aisaka , Haruko Hiraike , Hiroe Hyodo , Seiichiro Obata , Osamu Hiraike , Hironobu
2
Hyodo
1

Department of OB/GYN, Hamada Hospital, Tokyo, 2Department of OB/GYN, University of Tokyo, Japan

P-029

rFSH combined with uhMG had significantly increased pregnancy compared withrFSH alone undergoing ovarian stimulation following a long protocol in IVF
1

2

1

1

1

○K.H. Lee , J.D. Cho , H.G. Sun , I.H. Park , J.H. Lee , S.K. Kim
1

1

2

Mamapapa&baby OB&GY, Ellemedi Infertility Clinic, Changwon, Gyoung Nam, Repubilic of Korea

P-030

A comparison of pregnancy outcome in women with normal oocyte reserve
according to the use of high or low dose follicle stimulating hormone (FSH)
stimulation○Gabrielle Citrino, Jerome H Check, Jung K Choe
UMDNJ, Robert Wood Johnson Med. School at Camden, Cooper Hosp./Univ. Med. Cntr., Dept. OB/GYN, Div. Repro. Endo.
& Infertility, Camden, NJ, US

P-031

The effects of laser assisted hatching on pregnancy rates○Forouzan Absalan

1,21Anatomical department, Jundishapoor university of medical sciences, Ahvaz, , 2Iranshiraz infertility and limited surgicalcenter, Shiraz, Iran

P-032

Correlation between semen intracellular parameters and fertilization rates following
ICSI○Mina Sharbatoghli, Bita Ebrahimi, Mojtaba Rezazade Valojerdi
Embryology Department, Cell Sciences Research Center, Royan Institute, ACECR, Tehran, Iran

P-033

Whole ooplasmic transfer by direct injection method using Piezo driven system forprevention of mutated mitochondrial DNA transmission
1

1

1

1

1

○Akiko Yabuuchi , Noriko Kagawa , Kenji Ezoe , Chiemi Mori , Yuko Takayama ,
2
1
1
1
1
Masashige Kuwayama , Keiichi Kato , Fumihito Aono , Yuji Takehara , Osamu Kato
1

Advanced medical research institute of fertility, Kato Ladies Clinic, Shinjuku, Tokyo, Japan, 2Repro-support medical research
center, Shinjuku, Tokyo, apan

P-034

Elevation of the Risk of Monozygotic Twinning by Expanded Blastocyst Transfer○Kazusuke Nagoshi
Nagoshi Ladies Clinic, Okayama, Japan

P-035

Evaluation of sperm chromatin integrity following IMSI or HBA for pre-ICSI sperm
selection○Satoshi Ueno, Kazuo Uchiyama, Keiichi Kato, Yuji Takehara, Osamu Kato
Kato Ladies Clinic, Tokyo, Japan

54

SCIENTIFIC PROGRAM

P-036

Differential effects of urinary and recombinant chorionic gonadotropin on oxidative
stress responses in decidualizing human endometrial stromal cell.
○Takeshi Kajihara, Japarath Prechanich, Hideno Tochigi, Osamu Ishihara
Saitama Medical University, Saitama, Japan

P-037

Heparin prevents programmed cell death induced by oxidative stress in humandecidualized endometrial stromal cells
Takeshi Kajihara, ○Japarath Prechanich, Hideno Tochigi, Osamu Ishihara
Saitama Medical University, Saitama, Japan

P-038

The role of 5'AMP-activated protein kinase (AMPK) in human endometrial stromalcells○Yasushi Kawano, Sinya Karakida, Yufuko Utsunomiya, Hisashi Narahara
Oita University, Oita, Japan

P-039

Pinopode Formation upon Progesterone, hCG, and Clomiphene Citrate treatment
M Kabir-Salmani, ○M Shahali
National Institute of Genetic Engineering and Biotechnology, Dept. Molecular Genetics, Tehran. Iran

P-040

IS THERE ANY CORRELATION OF UTERUS POSITION AND DEEPTH EMBRYO
TRANSFER ON PREGNANCY RATE IN IVF○Agus Supriyadi
Harapankita hospital, Republic of Indonesia

P-041

Influence of antisperm antibodies (ASA) on the outcome of infertility treatment○Yasufumi Shimizu, Takeshi Yorimitsu, Hiroshi Motoyama, Motohiro Ohara,
Toshihiro Kawamura
Denentoshi Ladies Clinic, Kanagawa, Japan

P-042

The effect of ovarian stimulation on expression level of E-cadherin, Leukemia
inhibitory factor, Progesterone receptor and αvβ3 Integrin genes in mouse
blastocysts○Bahar Movaghar, Saeedeh Askarian
Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

P-043

Usefulness of elective single blastocyst transfer in the first treatment cycle forinfertile women aged less than 35.
○Hiroaki Shibahara, Shoko Hashimoto, Shiho Nagayama, Kazuhiko Shimada,
Tatsuya Suzuki, Mitsuaki Suzuki
Department of Obstetrics and Gynecology, School of Medicine, Jichi Medical University, Tochigi, Japan

P-044

A Summary of The clinical outcome of minimal stimulation IVF/ ICSIYan Ping Ma, Ze Wu, Bo Deng, ○Yonggang Li
Department of Reproduction and Genetics, Reproductive Medicine Centre, The First Peoples Hospital of Yunnan Province,
KunMing,China

P-045

Ovarian Response and Reserve Evaluation in consecutive IVF/ICSI-ET cycles○Mujdegul Karaca, A. Sertac Batioglu, Murat Ozel, Selcen Bahadir, Simla Karaca,
Beril Gurlek
Dr ZTB Women's Health Hospital, Ankara, Turkey

55

SCIENTIFIC PROGRAM

P-046

Our 11-year experience with sperm cryopreservation for patients with malignant
disease○Syuichi Iida, Masakuni Suzuki, Kazuhiro Hirayama, Ikuo Tachibana, Masahiro Katou,
Kouhei Tanaka
M. Suzuki's Memorial Hospital, Miyagi, Japan

P-047

A Successful pregnancy and live birth after intracytoplasmic sperm injection withcryopreserved limited numbers of spermatozoa stored in the practical container○Yuji Endo, Yoshitaka Fujii, Hiroaki Motoyama
Kurashiki Medical Clinic, Okayama, Japan

P-048

An evaluation of a modified slow cool cryopreservation technique and the efficacy ofa graduated estrogen regimen vs. natural cycles on pregnancy rates following frozen
embryo transfer○Jung. K Choe, Jerome H Check
UMDNJ, Robert Wood Johnson Med. School at Camden, Cooper Hosp./Univ. Med. Cntr., Dept OB/GYN, Div. Repro. Endo.
& Infertility, Camden, New Jersey, U.S.

P-049

The effect of vitrification on Histone Modification of regulatory regions of H19, Igf2
and Mest imprinted genes in mouse embryo
1

1

2

3

○Bahar Movaghar , Saeideh Sahraei , Maryam Shahhoseini , Ali Farrokhi
1

2

Department of Embryology, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran, Department of Genetics,
Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran, 3Department of Stem Cell, Royan Institute for
Reproductive Biomedicine, ACECR, Tehran, Iran

P-050

Withdrawn

P-051

Which is more effective for pregnancy freezing embryos: in day-2, day-3 stage or
blastocyst?
○Tomio Sawada, Kaori Yoshikai, Sayumi Hori
The sawada women's clinic Nagoya reproduction center, Aichi, Japan

P-052

Highest liquid nitrogen quality for vitrification process : Micro bacteriological filtrationof LN2
1

2

3

4

5

Ana Cobo , ○Damia Castello , B Weiss , C Vivier , Angel De La Macorra , F Kramp

6

1

IVI clinica de Fertitidad, Valencia, Spain, 2IVI clinica de Fertitidad, 3Air Liquide Head Office Industrial Merchant, 4Air
Liquide Sante International, 5Air Liquide Medicinal, 6Soprodi S.A.S., Quai du canal, Spain

P-053

Comparing developmental potential and ultra structure of human blastocysts after
re-vitrification
1

2

1,2

○Mansoureh Movahedin , Zahra Alamolhoda , Nasim Ghorbanmehr , Mojgan Moradkhani
1Tarbiat Modares University, Tehran, Iran, 2Shayamehr IVF Center, Tehran, Iran, 3Khatamolanbia Hospital, Tehran, Iran

P-054

Examination of blastocyst transfer after pronucleate embryos were slowly
freeze-thawed○Hiroei Ohhashi, Yukio Ohmomo, Osamu Arakawa
Arakawa Ohmomo Angel Mother Clinic, Niigata, Japan

56

3

SCIENTIFIC PROGRAM

P-055

5/-(N-Ethylcarboxamido) adenosine (NECA) improves angiogenesis in transplanted
human ovarian tissue: potential for fertility preservation
1

2

3,4

5

○Maryam Hormozi , Saeed Talebi , Amir Hassan Zarnani , Mahmood Jeddi-Tehrani ,62Ladan Hosseinigohari , Mohammad Mehdi Akhondi
1Biochemistry Department, lorestan university of medical sciences, Khorramabad,Iran, 2Reproductive Biotechnology ResearchCenter, Avicenna Research Institute, (ACECR), Tehran, Iran, 3Nanobiotechnology Research Center, Avicenna ResearchInstitute, (ACECR), Tehran, Iran, 4Immunology Research Center, Tehran University of Medical Sciences, Tehran, Iran,
5
Monoclonal Antibody Research Center, Avicenna Research Institute, (ACECR), Tehran, Iran, 6Biochemistry Department,
Tehran university of Medical Sciences, Tehran, Iran

P-056

Assessing risks of birth defects in ART babies○Coralia V. Stefanescu, Ion M Rusa, George Costea, Lenuta Malcea
Euromaterna Hospital, Constanta, Romania

P-057

Two cases of ovarian abscess after oocyte retrieval○Aki Oride, Haruhiko Kanasaki, Kohji Miyazaki
Shimane University School of Medicine, Shimane, Japan

P-058

An experience of an infertility treatment for a woman with benign metastasizing
leiomyoma○Tsuyoshi Hashiba, Toshio Hamatani, Naoaki Kuji, Kou Sueoka, Yasunori Yoshimura
Keio University School of Medicine, Tokyo, Japan

P-059

A Transgenic Mouse Model of Breast Cancer Using tissue specific MMTV Promoter
1,2

1

1

2

○Maryam Shahali , Ehsan Ranaei , Shams-Ara Mahdi , Javad Mowla , Maryam
13Kabir-Salmani , Karim Nayernia
1National Institute of genetic engineering and Biotechnology, Tehran, Iran & Tarbiat Modares University, Tehran, Iran,
Tarbiat Modares University, Tehran, Iran, 3GENOCELL CO.

2

P-060

Effect of genistein administration on intratesticular testosterone level andspermatogenesis in rats treated with busulfan○Heejun Chi, Kangwoo Cheon, Jonghyun kim, Giyoung Kim, Jaeseok Lee, Sungil Roh
i-Dream Center, Mizmedi Hospital, Seoul, Korea

P-061

The role of Doppler in predicting endometrial thickness, pattern and blood flow ininfertile women undergoing IVF cycle in a tertiary care institute○Anupama Bahadur, Neeta Singh, Neena Malhotra, Ashok Bhatt, Suneeta Mittal
All India Institute of Medical Sciences, New Delhi, India

P-062

Reduced blastcyst formation rate in women in advanced reproductive age with highAnti-Mullerian hormone level○Takahiro Noda, Shinako Hashimoto, Mitsuru Iwamoto, Syuichi Iida, Kohei Tanaka,
Masakuni Suzuki
Suzuki Memorial Hospital, Miyagi, Japan

P-063

Effects of salpingectomy on ovarian response in controlled ovarian hyperstimulation
for in vitro fertilization: a reappraisal
1○Israel Wagman , Benny Almog
2
Tulandi

1,2

1

1

1

, Ishai Levin , Gali Barkn , Dina Kovalsky , Togas

1

Tel Aviv Sourasky Medical Center, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel., 2Department of
Obstetrics and Gynecology, McGill University, Montreal, Quebec, Canada

57

SCIENTIFIC PROGRAM

P-064

Semen analysis and evaluation of ICSI outcome in patients infected with hepatitis B
virus
1

2

2

1

2

○Mounir Ajina , Neila Hannachi , Khaled Youssef , Souhir Mehri , Jalel Boukadida ,3
Ali Saad1

Unit of reproductive Medicine, University Hospitol Fahat Hached, Sousse, Tunisia, 2Laboratory of Bacteriology, University
Hospital Farhat.Hached, Sousse, Tunisia, 3Laboratories of Cytogenetic, Molecular Biology and Human Biology of
Reproduction, Farhat Hached Hospital, Sousse, Tunisia.

P-065

Effect of Chemotherapy on Spermatogenesis in Testicular Cancer Patients
1

1

1

2

1

○Miki Fuse , Takashi Imamoto , Takanobu Utsumi , Takumi Endo , Naoki Nihei ,1
Tomohiko Ichikawa
1

Department of Urology, Graduate School of Medicine, Chiba University, Chiba, Japan, 2Department of Urology, Toho
University Sakura Medical Center, Chiba, Japan

P-066

Reproductive outcomes following laparoscopic myomectomy in patients withinfertility○Kohei Tanaka, Masakuni Suzuki
M. Suzuki Memorial Hospital, Miyagi, Japan

P-067

The role of CINC/gro in the rat ovulatory process○Yu Tanaka, Akira Kuwahara, Yuya Yano, Yuri Yamamoto, Kenji Hinokio, Minoru Irahara
Deparatment of Obstetrics and Gynecology, The University of Tokushima, Institute for Health Biosciences, Tokushima, Japan

P-068

A Commercially Available Dual-Buffered IVF Handling Medium Containing HEPES
and MOPS Maintains Stable pH and Supports Human Sperm Survival, Normal
Fertilization Following ICSI and Embryo Development1○Jason E Swain , Marlane

2

3

4

Angel , Nadir Cira , Juergen Liebermann , Thomas Pool

5

1

University of Michigan, Ann Arbor, MI, 2Laurel Fertility Care, San Francisco, CA, USA, 3Bacchi IVF Center, Istanbul,
Turkey, 4Fertility Center of Illinois, Chicago, IL, USA, 5Fertility Center of San Antonio, San Antonio, TX, USA

P-069

Development of Mouse and Human Embryos in a Low Humidity Incubator○Chiemi Mori, Tadashi Okimura, Fumihito Aono, Yuji Takehara, Osamu Kato
Kato Ladies Clinic, Tokyo, Japan

P-070

Contribution of oocyte and embryo cleavage quality in obtaining blastocysts
1

1

1

2

2

○Mounir Ajina , Sonia Jallad , Souhir Mehri , Sawsen Meddeb , Hedi Khairi , Ali Saad

3

1

Unit of reproductive Medicine. University Hospital Farhat.Hached, Sousse, Tunisia, 2Department of Obstetrics andGynaecology, University hospital Farhat.Hached, Sousse, Tunisia., 3Laboratories of Cytogenetic, Molecular Biology and
Human Biology of Reproduction Farhat Hached Hospital, Sousse-Tunisia

P-071

Involvement of first cytokinesis on gene expression at blastocyst stage○Satoshi Sugimura, Tadayuki Yamanouchi, Yutaka Hashiyada, Eiji Kobayashi, Kei ImaiNational Livestock Breeding Center, Fukushima, Japan

P-072

Zona-free oocyte formed a one pronucleus zygote following piezo intracytoplasmic
sperm injection can subsequently develop to the blastocyst in micro-well culture
system. (Case Report)○Xinzhi Yang, Akina Takamura, Emi Fukunaga, Tomoyo Kusuda, Shinichiro Okano,
Masayuki Kinutani
Kinutani Women's Clinic, Hiroshima, Japan

P-073

A CUMULATIVE EMBRYO SCORING SYSTEM USED TO PREDICTPREGNANCY OUTCOME○Yoon Jeong Choi, Jung Ho Kim, Chan Park, Kwang Rae Kim, Sung il Roh, Hee Jun chi
i-Dream Center, Mizmedi Hospital, Seoul, Korea

58

SCIENTIFIC PROGRAM

P-074

Cytoplasmic halo as an additional indicator to predict the quality of the results of IVF
and ICSI embryos○Devi Natalia, Harris Harlianto, Sintya Jatnikasari, Tono Djuwantono, Wiryawan Permadi
Aster Clinic, RS.Hasan Sadikin, Bandung, Indonesia

P-075

Withdrawn

P-076

Effects of L/D stimuli and the circadian clock on reproductive physiology in female
mice
1

2

○Tomoko Amano , Juergen Ripperger , Urs Albrecht

2

1

Kinki University, Japan, 2Dep. of Medicine, Div. of Biochemistry, University of Fribourg

P-077

Numbers of CGG repeats on the FMR1 gene in Japanese patients with premature
ovarian insufficiency○Naoki Okamoto, Naomi Hamada, Yodo Sugishita, Nobuhito Yoshioka, Noriyuki Takahashi,
Bunpei Ishizuka
Department of Obstetrics and Gynecology, St.Marianna University School of Medicine

P-078

Genetic analysis of spermatozoa in four infertile patients with macrocephalic sperm
head syndrome
1,2

3

4

1

1

○Ghaya Merdassi , Pierre Ray , Meriem Chaabouni , Nedia Memmi , Fethi Zhioua , Amel
1
Zhioua1

IVF Center, Aziza Othmana Hopsital, Tunisia, 2Faculty of pharmacy.Monastir.Tunisia, 3Department of genetic andprocreaction. Tronche Hospital .grenoble. France , 4Department of human genetic. Charles Nicolle Hospital .Tunisia

P-079

Gene Expression Profiling of Karyotypically Normal, Trisomy 12 and XXY Human
Embryonic Stem Cells
1

1,2

2

2

1,2

Hye Won Seol , ○Sun Kyung Oh , Baik Seol Cho , Kyung Eui Park , Young Min Choi ,1,2Shin Yong Moon
1

Institute of Reproductive Medicine Population, Medical Research Center, Seoul National University College of Medicine,
Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul 110-799, Korea

2

P-080

Effect of dibutyryl cAMP during in vitro maturation on the developmental
competence of bovine oocytes after ICSI
1,2

2

2

2

○Chikako Kani , Tomohisa Wada , Akiko Kuwahata , Masanori Ochi , Toshitaka Horiuchi

11Graduate School of Comprehensive Scientific Research, Prefectural University of Hiroshima, Hiroshima 727-0023, Japan,
Ochi Yume Clinic Nagoya, Aichi, Japan

2

P-081

Androstenedione induces abnormalities in morphology and function of developing
oocytes, which impairs oocyte meiotic competence
1

1

1

1

2

○Wataru Tarumi , Sanae Tsukamoto , Yuki Okutsu , Noriyuki Takahashi , Masanori Itoh ,1
Bunpei Ishizuka
1

Department of Obstetrics and Gynecology, St. Marianna University School of Medicine, Kanagawa, Japan., 2Department of
Biology, College of Liberal Arts and Sciences, Tokyo Medical and Dental University, Ichikawa, Chiba 272-0827, Japan.

P-082

Preimplantation genetic diagnosis for spinal muscular atrophy usingmini-sequencing and genetic linkage analyses at the blastocyst stage
1,2

3

3,4

○Chia-Cheng Hung , Shee-Uan Chen , Shin-Yu Lin , Yi-Ning Su
1

1,2,4

Graduate Institute of Clinical Genomics, National Taiwan University College of Medicine, Taipei, Taiwan 2, Department ofMedical Genetics, National Taiwan University Hospital, Taipei, Taiwan, 3Department of Obstetrics and Gynecology, National
Taiwan University Hospital, Taipei, Taiwan, 4Graduate Institute of Clinical Medicine, National Taiwan University College ofMedicine, Taipei, Taiwan

59

SCIENTIFIC PROGRAM

P-083

Preimplantation Genetic Diagnosis (PGD) for translocation carrier with mild
stimulation and single vitrified blastocyst transfer : Case Report○Naoki Aoyama,Yuji Takehara, Satoshi Kawachiya,Tomoko Kuroda, Nami Kawasaki,
Rie Yamadera,Keiichi Kato, Osamu Kato
Kato Ladies Clinic, Tokyo, Japan

P-084

Noninvasive morphological examination for germinal vesicle oocytes predict theirmaturity process of completion
1

1

1

1

○Tsuyoshi Okubo , Ryoko Matsumoto , Naoko Shimada , Teruaki Hayashi ,1
1
2
Tomoya Segawa , Shoukichi Teramoto , Masashige Kuwayama
1

Shimbashi Yume Clinic, Tokyo, Japan, 2Repro-Support Medical Research Center, Tokyo, JapanP-085

hCG is critical for the uterine decidual response in mice
1

1

1

1

2

○Hiroomi Kawano , Kenji Ezoe , Noriko Kagawa , Akiko Yabuuchi , Keiko Ochiai ,, Hiroshi
2
1
1
1
1
Nagashima ,, Hisao Osada , Fumihito Aono ,Yuji Takehara , Osamu Kato
1

Kato Ladies Clinic, Tokyo, Japan 2Meiji University, Kanagawa, Japan

P-086

Cytogenetic study in early spontaneous abortion after IVF and ICSI○Satoshi Kawachiya, Yuji Takehara, Keiichi Kato, Hisao Osada, Naoki Aoyama, Osamu Kato
Kato Ladies Clinic, Tokyo, Japan

P-087

Day 7 -hCG level after frozen-thawed blastocyst transfer can be a predictor for
pregnancy outcomes?
1

2

1

1

1

○Keiichi Kato , Tomoya Segawa , Hisao Osada , Tamotsu Kobayashi , Yuji Takehara ,1
Osamu Kato
1

Kato Ladies Clinic, 2Shimbashi Yume Clinic, Tokyo, Japan

11. Luncheon Seminars:
Luncheon Seminar 1
Sunday, September 11 12:00～13:00 (Concord Ballroom A)
Chairperson: Takafumi Utsunomiya (Japan)
12:00～13:00
GnRH antagonists in ovarian stimulation for IVF and ICSI frompractical perspective-Atsushi Tanaka (Japan)
Luncheon Seminar 2
Sunday, September 11 12:00～13:00 (Concord Ballroom B)
Chairperson: Osamu Ishihara (Japan)
12:00～13:00
Clinical aspects of GnRH analogue use in Assisted reproduction-Naoaki Kuji (Japan)
Luncheon Seminar 3: Potential of Piezo-assisted micromanipulation brought by PMM from thefundamental research to the clinical application
Sunday, September 11 12:00～13:00 (Concord Ballroom C)
Chairperson: Fumihito Aono (Japan)
12:00～12:20
Impact of piezo impact drive unit; it reversed the common sense oflife-Teruhiko Wakayama (Japan)
12:20～12:40

Would you try Piezo -ICSI without mercury?
-Hiroshi Morita (Japan)

60

SCIENTIFIC PROGRAM

12:40～13:00

Advantage of the use of Piezo driven system (PMM) in ARTs;
Whole ooplasmic replacement in Germinal Vesicle oocytes-Akiko Yabuuchi (Japan)

Luncheon Seminar 4
Monday, September 12 12:00～13:00 (Concord Ballroom A)
Chairperson: Yoshiharu Morimoto (Japan)
12:00～13:00
The role of AMH in female reproduction-Budi Wiweko (Indonesia)
Luncheon Seminar 5
Monday, September 12 12:00～13:00 (Concord Ballroom C)
Chairperson: Yuji Takehara (Japan)
12:00～13:00
Fertility tourism in the U.S.
-John Zhang (USA)
Luncheon Seminar 6
Tuesday, September 13 12:00～13:00 (Concord Ballroom A)
Chairperson: Osamu Ishihara (Japan)
12:00～13:00
Global inequity: Access to Assisted Reproductive Technologies-Sheryl Vanderpoel (Switzerland)61

SPECIAL GUEST LECTURE

62

●Special Guest Lecture

Induction of Pluripotency by Defined Factors
Shinya Yamanaka
Director, Center for iPS Cell Research and Application (CiRA),
Professor, Institute for Integrated Cell-Material Sciences (iCeMS),
Kyoto University
Induced pluripotent stem (iPS) cells were originally generated from mouse and human fibroblasts byretroviral introduction of Oct3/4, Sox2, c-Myc, and Klf4. iPS cells are similar to embryonic stem (ES) cells inmorphology, proliferation, gene expression, and most importantly, pluripotency. Compared to ES cells, iPS
cells have less ethical controversy and can be generated from various genetically identified individuals
including disease patients or those having specific human leukocyte antigen (HLA) types. Patient-specific iPS
cells provide unprecedented opportunities in disease research, drug screening, and toxicology. A bank of iPS
cells constructed from HLA-homozygous donors would provide significant resources for stem cell therapy.
However, recent reports of tumor formation following transplantation, and the large diversity between iPS cellclones highlight potential problems. Furthermore, the mechanism of reprogramming remains unclear.
In addition to fibroblasts, iPS cells can be generated from various somatic cells, such as hepatic cells,
gastric epithelial cells, neural cells, dental pulp cells, peripheral blood cells, and cord blood cells. As
alternatives to retroviral transduction, iPS cells can be generated by lentiviruses, adenoviruses, plasmids,
transposons, recombinant proteins, or synthesized mRNA. Recently, we reported an integration-free induction
method using episomal vectors. This method can induce human iPS cells efficiently and reproducibly.
Regarding iPS cell induction factors, we discovered that L-Myc and the transcription factor Glis1, which isstrongly expressed in the unfertilized egg, can establish iPS cells with a high efficiency and quality, replacing
the oncogene c-Myc. Other reports suggest that chemicals can further enhance induction efficiency.
Each induction experiment can result in up to 100 or more independent iPS cell clones. These iPS cellclones may vary qualitatively, considering responses to in vitro directed differentiation protocols and theirpropensity to produce tumors. In fact, we have previously shown that the origins of mouse iPS cells haveprofound effects on tumorigenicity. It is therefore essential to determine the best origins, the best induction
protocols, and the best methods to evaluate iPS cell clones and subclones for future clinical applications. From
this point of view, the need for genetic and epigenetic analyses, such as DNA methylation, histone
modification, and genomic imprinting becomes more significant. It is also important to note that iPS cellwithin a clone can be heterogeneous, despite their common derivation from a single progenitor cell. This is
likely because the process requires multiple cell division and cannot be completed by the four exogenous
factors alone. Additional endogenous factors are required to achieve full reprogramming. Better understandingof the reprogramming mechanism will facilitate more uniform and complete reprogramming during iPS cell
generation.
◆Biosketch
Titles: Director, Center for iPS Cell Research and Application (CiRA), Kyoto University
Professor, Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University

63

●Special Guest Lecture

Educational History:
1981–1987

Kobe University, Kobe, Japan
School of Medicine (M.D. awarded in March, 1987)

1989–1993

Osaka City University Graduate School, Osaka, Japan
Division of Medicine (Ph.D. awarded in March, 1993)

Professional History:
1987–1989

Resident, National Osaka Hospital, Osaka, Japan

1993–1995

Postdoctoral Fellow, Gladstone Institute of Cardiovascular Disease
The J. David Gladstone Institutes, San Francisco, CA, USA

1995–1996

Staff Research Investigator, Gladstone Institute of Cardiovascular Disease
The J. David Gladstone Institutes, San Francisco, CA, USA

1996–1999

Assistant Professor, Osaka City University, Medical School, Osaka, Japan

1999–2003

Associate Professor, Nara Institute of Science and Technology, Nara, Japan

2003–2005

Professor, Nara Institute of Science and Technology, Nara, Japan

2004–

Professor, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan

2007–

Professor of Anatomy, University of California, San Francisco, CA, USA

2007–

Senior Investigator, Gladstone Institute of Cardiovascular Disease
The J. David Gladstone Institutes, San Francisco, CA USA

2007–

Professor, Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto,
Japan

2008–

Director, Center for iPS Cell Research and Application (CiRA), iCeMS,
Kyoto University, Kyoto, Japan2010–

Director, Center for iPS Cell Research and Application (CiRA), Kyoto University,
Kyoto, Japan

Honors and Awards:
2007
2008
2008
2008
2009
2009
2010
2010
2010
2010
2011
2011
2011

JSPS Prize, Japan Society for the Promotion of Science
The Special Prize for Science and Technology, the Minister of Education, Culture, Sports, Scienceand Technology, JapanThe Shaw Prize in Life Science and Medicine, Hong Kong
Robert-Koch Preis 2008, Germany
Albert Lasker Basic Medical Research Award, USA
Canada Gairdner International Award, The Gairdner Foundation, Canada
100th Imperial Prize and Japan Academy Prize, The Japan Academy, Japan
Selected as Person of Cultural Merit (―Bunka Korosha‖), Japan
26th annual Kyoto Prize in Advanced Technology, Japan
Balzan Prize for Stem Cells: Biology and Potential Applications, Italy
King Faisal International Prize, Saudi Arabia
Elected as National Academy of Sciences Members and Foreign Associates, USA
Wolf Prize in Medicine, Israel
etc.

64

PRE-CONGRESSWORKSHOPS
Vitrification
Pre-implantation Genetic Diagnosis

65

●Pre-Congress Workshop: Vitrification 1

Basic science of vitrification
Stanley P. Leibo
Department of Biological Sciences, University of New Orleans,
New Orleans, LA, U.S.A.
AIMS: The purpose of this lecture is to briefly review basic principles ofcryopreservation as applied to vitrification.
METHODS: Over the past decade, ~800 articles have described cryopreservation of oocytes and embryos by―vitrification‖, the reversible transition of liquid into an amorphous non-crystalline glass. Vitrification
presumably requires high concentrations of cryoprotective additives (CPAs) and very high cooling rates to
produce the glassy state. Vitrification of reproductive cells and tissues has been achieved by suspending themin solutions containing 15 to 30% permeating CPAs plus 18% saccharide, and cooling very small samples at
rates ≥10,000°C/min. One criterion considered to be essential if the best results are to be achieved is that theextracellular solution itself must form a glass when cooled at high rates.
RESULTS: Many experimental and clinical studies have been published in which survival rates of vitrified
samples ranging from 75% to close to 100% have been reported. Comparison of results attained byvitrification with results from standard, equilibrium cooling methods have almost always demonstrated thatvitrification is as good and usually better than standard freezing. One limitation of many vitrification methods
is that human specimens, to be cooled at high rates, are immersed directly into liquid nitrogen. This may
expose them to potential contamination by viruses or microbes.
CONCLUSIONS: Recent results and re-examination of older results reveal that it may not be necessary tovitrify the extracellular solution to cryopreserve oocytes and embryos. The key to survival is whether the
intracellular contents vitrify or not. Ultimate survival of cryopreserved cells and tissues may be more
dependent on the rate at which the specimen is warmed, rather than the rate at which it is cooled.
◆Biosketch
Stanley P. Leibo received his Bachelor of Arts degree from Brown University, Master of Science degree fromthe University of Vermont, and Master of Arts and Doctor of Philosophy degree in Biology from Princeton
University. He began his career in cryobiology with Peter Mazur at Oak Ridge National Laboratory in
Tennessee. He then became Vice-President of Research and Development at Rio Vista International, a cattlecompany in San Antonio, Texas. In 1988, he was appointed Associate Professor of Obstetrics and Gynecology
and of Urology at Baylor College of Medicine in Houston. From 1991 to 1998, Dr. Leibo was Professor of
Biomedical Sciences at the Ontario Veterinary College of the University of Guelph in Ontario, Canada. In
1998, he was named to his present positions as Professor of Biological Sciences at the University of New
Orleans, and Senior Scientist at the Audubon Center for Research of Endangered Species in New Orleans.
In addition to his permanent positions, Dr. Leibo has served as a Fellow of the Japan Society for the
Promotion of Science at Kyoto University, and for four years was Visiting Professor on the medical faculty at
the Dutch-speaking Free University in Brussels, Belgium. He was elected president both of the Society for
Cryobiology and of the International Embryo Transfer Society, and was named an Honorary Lifetime Memberof the American Embryo Transfer Association. In 2009, Leibo received the Pioneer Award of the International
Embryo Transfer Society to recognize his role in the derivation of the original methods to cryopreserve
gametes and embryos. He has published more than 135 scientific articles and book chapters, and has delivered
more than 180 lectures in the United States and in twenty-nine other countries.

66

●Pre-Congress Workshop: Vitrification 2

Ovarian reserving
Noriko Kagawa
Kato Ladies Clinic, Tokyo, Japan
Recently, the adaptation of the cryopreservation has been expanding and dramaticallychanging in human assisted reproduction field because of the development of the ultra
rapid vitrification method. This technique enables to establish oocyte bank which preserve fertility of cancer
women after the therapy by combination with IVF cycle. To preserve fertility in female cancer patients,
oocyte vitrification is one solution. However, it does not work for the children and the urgent patients who do
not have enough time for one or more IVF cycles. Ovarian tissue cryopreservation has the potential to solvethese problems, and to preserve their natural fertility after chemo and radiotherapy. Similarly to oocyte
freezing, ovarian tissue cryopreservation has utilized a conventional slow freezing method, resulting in limited
success. However, encouraging good results with ovarian tissue freezing are recently being obtained by using
vitrification. More recently, we also established the efficient ovarian tissue cryopreservation by ultra rapid
vitrification, the Cryotissue method, and already applied for the young unmarried cancer patient for the firsttime in the world. So we had have also the first success of vitrified ovarian tissue transplantation between
sisters who HLA matched. She had the first period in April 2008 after ovarian tissue transplantation since she
finished cancer therapy in 2004. And Second success of vitrified ovarian tissue transplantation between
identical twins in February 2010. She had the first increasing of estradiol and decreasing of FSH and LH at
42days after ovarian tissue transplantation. This ovarian tissue vitrification can preserve sex of female patients
after cancer therapy, and help to improve their quality of life to be happy as women. In this lecture, I introduce
our detailed protocols, experimental/clinical results of ultra rapid vitrification for human ovarian tissue.
◆Biosketch
Dr. Noriko Kagawa obtained her Ph.D. in 2005 from the Kyoto University, Japan. She is currently the seniorscientist at the Advanced Medical Research Institute of Fertility in Kato Ladies' Clinic, Tokyo, Japan, theworld's largest human IVF unit. In 2000, she started animal embryology and studied Assisted Reproductive
Technologies, for example, IVM, IVF, vitrification, embryo culture in pig, and established the porcine
follicular growth system using SCID mice. She moved to the human IVF field in 2004, and has developed
vitrification methods for mammalian ovaries and ovarian tissues in mice, cat, dog, lion, giraffe, bovine, human.
Her major research interests have focused on in vitro growth and in vitro maturation for preantral follicles ofadult mammals as well as on vitrification of Germinal Vesicle stage oocytes and various organs for unlucky
patients.

67

●Pre-Congress Workshop: Vitrification 3

Ovary Transplant: Ovary allotransplantation between
non-homozygous sisters: Success and failure, an immunologic puzzle
Sherman J. Silber
Infertility Center of St. Louis
Objective: To see whether prior bone marrow transplantation from an allogeneic
ovary donor can allow an ovary cortical graft from the same donor to be successful without
immunosuppression.
Design: Two cases of ovary donation from a previous allogeneic bone marrow donor to her sister who became
sterile as a result.
Materials & Methods: In two pairs of non-identical, non-twin, sisters, leukemia was treated in the younger
sister (one at 4 years of age and one at 28 years of age) with total body irradiation, ablative chemotherapy, andbone marrow transplant. Both sisters were deemed cured by their oncologists, and both underwent ovarian
cortical transplant from their sister who had previously donated bone marrow.
Results: In both cases, prior to transplantation, DNA fingerprinting of tissue and peripheral lymphocytes
indicated complete engraftment of donor bone marrow. Yet in one case, whose cancer treatment and bonemarrow transplant was 3 years earlier, rejection of the graft occurred, and rejection of a later frozen cortical
graft also occurred. In the other case, menstruation and ovulation resumed at 63 days, and there was no
rejection, and the graft continues to function perfectly with no immunosuppression. In this case, HLA typing
had shown a perfect match prior to the transplant. In the other case, which surprisingly rejected, the HLA
match was not perfect before the bone marrow transplant, and the recipient had an increase in eosinophil
count, indicating a mild "graft versus host" condition (GVH).
Conclusions: Successful bone marrow transplant recipients thought to have complete engraftment of donorbone marrow might still have undetectable, clones of their original bone marrow that could mount a rejection
against an ovary cortical graft.
◆Biosketch
Dr. Sherman Silber is one of the world's leading authorities on IVF, microsurgery, vasectomy and tubal
reversal, egg and embryo freezing, ovary and testis transplantation, and the reproductive biological clock. He
is director of the Infertility Center of St. Louis at St. Luke's Hospital in St. Louis, and author of thebest-selling How To Get Pregnant.

68

●Pre-Congress Workshop: Vitrification 4

The Cryotop Methods (Live Demonstration)
Masashige Kuwayama
Repro-Support Medical Research Center, Tokyo, Japan
Recent drastic advances in cryobiology have made it possible to preserve various typesof reproductive cells with little viability loss. Vitrification, the alternative
cryopreservation method is a powerful tool to any biological specimens, which cannot be preserved by theconventional slow freezing. The Cryotop method, effective and safety ultra-rapid vitrification realized thesuccessful clinical use of vitrification not only for human PN zygotes, cleavage stage embryos and blastocysts
but also for oocytes. The Cryotop method has been clinically applied to more than 500,000 clinical cases forhuman oocytes and embryos for these 10 years in 40 countries with excellent results without any serious
problems like virus contamination during storage. The reasons of the advantage of Cryotop method is 1)
Reliable 2) Universal and 3) Simple. In this session, I show these advantages of the Cryotop method by a live
demonstration of the standard Cryotop method, and other 2 variations of the Cryotop methods adapted to theregulations in different countries.
1. Cryotop method for human oocytes (Standard version)
2. Cryotop method with Cold Air Sealing (Open cooling/Closed storage)
3. Cryotop method with Closed Cooling (Closed Top method)◆Biosketch
Name: Masashige Kuwayama Repro-Support Medical Research Center
Ph.D.: Hokkaido University
Occupation:
Present - CEO, Repro-Support MRC
2010- General Academic Supervisor, Kato Ladies group2007- Board Internal Society of MAAR
2005- Editor, J. Reproductive Bio Medicine Online
2005- Board, JSClE
2005- Lecturer, IUMW, Meiji, Azabu Univ.
Publications: 81 Papers on Scientific Journal: JRF, Reproduction, Cryobiology, CryoLetters, BOR etc..
290 Papers on Scientific Meeting: IETS, ICAR, ASRM, ESHRE etc.
Topics: Scientific Director, largest IVF center for 10 years.
R & D Director: vitrification, GVT, spermatogenesis, ES-cells.
Obtained first calf after vitrification of blastocyst (1991), MII oocyte (1992). First vitrification porcine
pre-hatch blastocyst, oocyte (1997, 1998). Developed Cryotop method for human oocyte (1999), embryo(2000), first oocytes bank (2001), in USA (2003). Vitrified 500,000 oocytes and embryos in 40 countries by
Cryotop method.

69

●Pre-Congress Workshop: PGD 1

Overview
Kou Sueoka
Keio University School of Medicine, Tokyo, Japan
Preimplantation genetic diagnosis (PGD) is going into the new era according to thedramatically developed technologies of genetic analysis. It is a medical approach tosupport transmission of genes to be hopefully healthy, which is the aim and the principle of reproductive
medicine. On the other hand, there has been a lot of ethical arguments to relate to criticism that it is medical
treatment to lead to selection of life including on the aspects of the indication and its range. The situation of
PGD is still different in the countries. There are the countries which are not accepted at all, while there arepositive countries approving it legally. Japan is a country that asks strict correspondence to require double
ethical deliberation and approval on the restrictive acclimated indications and also the diagnostic method.
ESHRE PGD Consortium has been collected the accumulated data on PGD outcome from the cooperated
institutions in the world and reported for a fixed interval. As a technical aspect of PGD the large changes haveoccured both in the alteration of biopsy and genetic diagnosis. However, biopsy method of next generation
PGD is shifting to that from polar body and blastocyst. Polar body biopsy is useful to distinguish mosaicism
of numerous anomalies. In addition, the method of blastocyst biopsy is also another alteration to improve
diagnostic accuracy by obtaining many cells, and to cope with wide variety of genetic diagnosis. As for thetechnologies for genetic diagnosis, while FISH has been conventionally used for diagnosis of chromosomal
information, nested PCR has been used for diagnosis of DNA information. However, the technical limitations
and problems have been experienced in both diagnostic methods so far. However, particularly diagnostic
significance of aneuploidy screening using FISH for limited number of chromosomes has been asked from thestandpoint of the evidence, and desired to shift to the new analytical technologies. Moreover, FISH is given asa basic problem in diagnostic accuracy from single cell based on the extremely restrictive information onlywith a number of detected signals. On the other hand, a lot of problems about PCR used for DNA diagnosis
has been noticed; for example, that tailor-made diagnostic method needs to be suffered in most cases
corresponding to a wide variety of genetic type; problems relating to gene amplification such as DNA
contamination and allele drop out; that there is limitation of DNA amount to diagnose for single copy gene of
single cell; and the subject for the cost and spending time. Genetic diagnosis is going into the new era ofgenome wide analysis. Array technology has been developed as a new trend of genetic diagnosis, and themethod of whole genome amplification (WGA), which is required prior to the array analysis for single cell,has been introduced into PGD. These technologies would be reaching the possibilities not only of largely
expanded diagnostic width, but also of improvement of diagnostic accuracy. However, from the viewpoint ofgenetic information management, we should pay more attention to perform enough genetic counseling for aclient.
◆Biosketch
Birth Date:
Present Status:

July 7, 1954.
Keio University School of Medicine, Department of Obstetrics and Gynecology. Associate
Professor, Director of Reproductive. Obstetrics and Gynecology.
Subspecialty:
Reproductive medicine, Clinical human genetics, Gynecologic endoscope surgery.
Title:
National license of medical doctor of Japan, Doctor of Medical Science (Ph.D.),
Specialty Board: Obstetricians and Gynecologist, Endoscopy, Clinical Genetics, Reproductive Medicine.
Career of Medical Background:1980.8
Graduation of Keio University School of Medicine,70

●Pre-Congress Workshop: PGD 1

1983.6
Keio University fellow,
1986.7
Johns Hopkins University post doctoral fellow (U.S.A., Maryland),
2001.11 Director of Genetic Counseling Section Keio University Hospital,
2000.4
Keio University Associate Professor.
Major Appointments: Vice president of the Alumni Society of Keio University School of Medicine.

71

●Pre-Congress Workshop: PGD 2Whole genome amplification is a new basic technology fornext-generation preimplantation genetic diagnosis on mendelian
inheritance disease
Suguru Sato, Kou Sueoka, Akira Nakabayashi, Yasunori Yoshimura
Department of Obstetrics and Gynecology, Keio University School of
Medicine, Tokyo, Japan
Preimplantation genetic diagnosis (PGD) is an established procedure of embryo genetic analysis, allows
couples carrying disorder to have an unaffected child, without invasive prenatal diagnosis and termination ofpregnancy. The scarcity of DNA is a limiting factor in many reseach fields. For PGD on Mendelian
inheritance diseases, because of difficulty in analyzing target single gene from a single cell, two rounds of
PCR, termed nested PCR is once a essential basic technique. Genetic diagnosis has been made by testing PCR
products for a variety genetic disorder. It requires appropriate primers based on genetic information derived
from the client's family and optimization of multiplex PCR condition and primer setting are labor-intensive.
There are limits to multiplexing that multilocus simultaneous analysis from a single cell is restrictive. In caseswith multiple exon deletions or more than two copies of mutation with autosomal gene, the contamination riskincreases. Development of new gene amplification method instead of PCR is desirable. Whole genome
amplification (WGA) is expected to be a solution for it. Multiple displacement amplification (MDA) is a
non-PCR based WGA which includes the bacteriophage φ 29 DNA polymerase, random hexamers expected
to allow multilocus parallel analysis by any combination of conventional genetic testing. For genetic diagnosis
of multiple loci, when one round WGA can amplify the template DNA instead of each 1st PCR on therelevant mutation site, risk of contamination reduces, leads time and cost reduction. One of the potential
problem associated with WGA is so called allele drop out (ADO), defined as the random non-amplification of
one of the alleles present in a heterozygous sample. To avoid that phenomenon, optimization of cell-lysis isnecessary. Method for cell-lysis using alkaline lysis, proteinase K, and other enzymes or detergents have been
expected for more efficient DNA collection. The combination of MDA and alkaline lysis extraction on single
cells by KOH has been highest efficiency for the amplification success rate from the target part from single
cell, which was stably more than 70%. Repetitive assay for mutation sites is favorable in PGD to avoid
misdiagnosis by ambiguous result especially for autosomal dominant diseases and compound heterozygote.
One of these diseases is Fukuyama congenital muscular dystrophy (FCMD), which is an autosomal recessive
disorder indigenous to Japan. The incidence of FCMD is 0.7-1.2 per 10,000 births, next to Duchenne muscular
dystrophy. Fukutin gene, responsible for FCMD is located on 9q31 and it is well known that almost all
patients have the 3-kb retrotransposal insertion in Fukutin gene and the disease often occurs as a result fromcombined homozygous for the mutation or compound heterozygous with other copy of point mutation. MDA
amplify scarce DNA enough to be served for multilocus simultaneous assay and abundant WGA products canbe analyzed repeatedly. Unbiased and high fedility WGA is an essential technique for the construction of
accurate and high through-put diagnostic system, will enable comprehensive mutation analysis bycombination of array-based technology. Further validation will be needed for PGD in next-generation.
◆Biosketch
2002 Graduated from Iwate Medical University School of Medicine
2002 Residents of Obstetrics and Gynecology, Keio University School of Medicine
2004 Obstetrics and Gynecology, Toyoshina Red Cross Hospital2005 Obstetrics and Gynecology, Otawara Red Cross Hospital
2006 Research Associate, Keio University School of Medicine, Obstetrics and Gynecology
2010 Research Associate, Tokyo Dental College Ichikawa General Hospital, Obstetrics and Gynecology
2011 Research Associate, Keio University School of Medicine, Obstetrics and Gynecology

72

●Pre-Congress Workshop: PGD 3

To PGD or not to PGD? How many oocytes are needed to start PGD
with?
Ilan Tur-Kaspa
Institute for Human Reproduction (IHR), the Department of Ob/Gyn,
The University of Chicago, Chicago, IL, USA
Introduction: The optimal number of retrieved oocytes for successful IVF outcome depends primarily on
women's age and the ovarian stimulation protocol. PGD results significantly and independently of embryo
development reduce the number of embryos available for transfer, by 25 - 88 percent, depending on the typeand number of PGD tests performed. We investigated if there is a magic number of oocytes to start PGD with.
This information is valuable for professionals who consul women on ovarian stimulation and possible
outcome of IVF with or for PGD.
Material and Methods: In addition to literature review, outcome data from personal clinical experience with
562 consecutive IVF-PGD cycles in 4 years (168 cycles for 51 single gene disorders, 99 cycles for HLA
matching, 44 cycles for translocations, and 251 PGD cycles for aneuploidy) were analyzed by women's age,
number of oocytes retrieved, and type of PGD performed. Patient's ovarian reserve was estimated before
treatment by age, antral follicle count, and day-3 FSH levels, and accordingly changes in the stimulation
protocol or in medication dosage to try to safely reach oocyte yield of 10-15, were implemented.
Results: An increase in the number of oocytes retrieved improved the patients chance to have an ET withnormal embryos, as well as to have surplus blastocysts cryopreserved. Responding poorly to stimulation may
result from an inadequate stimulation protocol or because of low ovarian reserve, which may or may not be
related to the genetic disorder itself. Most young patients may have an ET with good outcome even with only
1-7 oocytes. However, for patients at age 35-42 years old, more oocytes significantly improved outcome.
Conclusions: Individually adjusted ovarian stimulation based on estimated ovarian reserve for patients
undergoing IVF-PGD improves outcome. When the patient is a poor responder, there is practically no risk of
OHSS and therefore, a mild stimulation protocol should not be offered. Once patients were adequately
stimulated and wish to conceive with their own eggs, oocyte retrieval and PGD may be continued even withvery low number (1-7) of oocytes or embryos. Routine canceling of PGD cycles because of poor response
should be reconsidered, especially for young women.
◆Biosketch
Professor Ilan Tur-Kaspa is the President and Medical Director of the Institute for Human Reproduction (IHR),
Director of the Clinical IVF-PGD Program, Reproductive Genetics Institute, and Professor at the Dept of
OB-GYN at the University of Chicago. Prof. Tur-Kaspa is one of the world's most experienced Reproductive
Specialists involved in advanced IVF-ICSI treatment with PGD. He specializes in advanced ART,
reproductive imaging, and PGD. He is the author and co-author of over 90 scientific publications and bookchapters. He serves as a Reviewer for the leading Journals in Reproductive Medicine, such as Fertility and
Sterility, Human Reproduction Update and Reproductive BioMedicine Online. He is a co-Editor of the bookon Biotechnology of Human Reproduction. He was awarded the 2010 Star Award of the American Society for
Reproductive Medicine (ASRM) for over 10 years of continuous scientific contributions in the field ofReproductive Medicine.

73

●Pre-Congress Workshop: PGD 4Introduction to performing efficient FISH-based PGD
Naoki Aoyama
Kato Ladies Clinic, Tokyo, Japan
Since Alan Handyside first established preimplantation genetic diagnosis (PGD)
more than 20 years ago, it has become a widely accepted clinical techniqueavailable in many centers around the world. Currently PGD‘s indication iswell-established for couples at high-risk for having children affected by hereditary diseases, habitual abortion,etc. Recently, microarray-based techniques have also been developed for the screening applied to the general
population. Nonetheless the ―classical‖ FISH technique is still an extremely useful, cost-effective andpowerful tool for PGD, especially for chromosomal translocation carriers.
The present workshop will provide you with an introduction to the fundamental techniques necessary forpracticing FISH-based PGD. Hopefully it will be a valuable aid to everybody wanting to start a PGD programat his/her center.
・Embryo biopsy
PGD implies the removal of polar bodies, blastomeres or trophectoderm cells from oocytes, cleavage-stageembryos or blastocysts, respectively. Embryo biopsy performed at cleavage-stage by the mechanical openingof the zona pellucida and suction of a single blastomere is considered the most fundamental technique and will
be demonstrated at the present workshop.
・Blastomere fixation
Blastomere fixation is one of the most critical steps in FISH-based PGD. Three methods for blastomere
fixation or spreading have already been described and currently all of them are considered acceptable.
Carnoy method : Methanol/Acetic acid
Cell spreading : Tween/HCl
Combined : Tween/HCl and Methanol/Acetic acid
Although the Combined method is the simplest and the easiest to perform, it was reported that the Carnoy
method might generate better nuclear quality by obtaining a wider nuclear diameter after fixation on a
glass-slide. Our own experience also supports the higher efficiency of the Carnoy method. An introduction
will be given to the three above-mentioned techniques highlighting the differences between each of them andhow to use them correctly to obtain accurate and reliable PGD results.
◆Biosketch
Major: Cytogenetics & Animal Physiology
Naoki Aoyama began his studies in biology in 1988 at Faculty of Animal Science University of Meij (Japan).
In 1994 he obtained his MSc degree in Agricultural Faculty from the same University.
After graduation he began working at Mitsubishi Biomedical Laboratories Co., Ltd and moved to Kato Ladies
Clinic in 2000.
He started to study Cytogenetics at Advanced Institute of Fertility in 2001 and developed his technique forPreimplantation Genetic Diagnosis.

74

PLENARY LECTURES

75

●Plenary Lecture 1

Natural/Clomiphene Cycle IVF: The Japanese Experience
Osamu Kato, M.D.
General Director, Kato Ladies Clinic, Shinjuku, Tokyo, Japan
During the some thirty years since Steptoe and Edwards delivered Louise Brown in
1978, in vitro fertilization and embryo transfer (IVF-ET) have come to be practiced allover the world. At the same time, Assisted Reproductive Technology has continued to make progress by leaps
and bounds with brilliant results, and various revolutionary techniques have been developed, solving eventhose problems which had been thought impossible to remedy. These techniques include Microsurgical
Epididymal Sperm Aspiration (MESA) (R. H. Asch and S. J. Silber, 1991), Intracytoplasmic Sperm Injection(ICSI) (Palermo et al., 1992), Percutaneous Epidydimal Sperm Aspiration (PESA) (O. Kato et al., 1993), andTesticle Sperm Extraction (TESE) (R. Schoysman et al., 1993) for male infertility; the Towako Method(transvaginal-transmyometrial embryo transfer for difficult transcervical embryo transfer cases) (O. Kato and
R. H. Asch, 1993), Pre-implantation Genetic Diagnosis (PGD) for screening embryos with abnormal genes,
and vitrification of unfertilized embryos for unmarried young cancer patients (M. Kuwayama, 2001) for
female infertility. However, as regards the ovulation stimulation method, protocols for Controlled OvarianHyperstimulation (COH) have not improved greatly, save for the use of different drugs. As a result, there haveemerged various questions, specifically:(1) Why do we retrieve so many oocytes despite the fact that only 2-3 oocytes are of good quality evenwhen more than ten oocytes are retrieved?
(2) Why do we need to transfer more than one embryo, increasing the multiple birth rate, when patients do
not wish to have more than one child? (Is it because it is difficult to select good quality oocytes or because
we want to improve the pregnancy rate?)(3) Why do we use COH which requires as the maturation trigger the use of HCG which has the potential
to induce severe Ovarian Hyperstimulation Syndrome (OHSS)?
(4) Why are we increasing the financial burden on patients with the use of drugs?
In our clinic, in order to address these questions, we have developed and practiced unique protocols for oocyte
retrieval and embryo transfer. Today, I would like to talk about the history of our protocols, from our first
Minimal Stimulation protocol up to our present protocol, the single follicle-single embryo transfer (SF-SET),
which requires no ovulation inducing drugs at all.
◆Biosketch
Osamu Kato, M.D., General Director, Kato Ladies Clinic
Born in1946 and graduated from Kanazawa University, School of Medicine in March 1972.
Medical Doctor in December 1972.
President, The International Association of Private Assisted Reproductive Technology
Vice-President, The World Association of Reproductive Medicine (WARM)
Executive Board Member, Japan Society of Fertilization and Implantation(JSFI)
Trustee, The International Society for Mild Approaches in Assisted Reproduction (ISMAAR)
Congress President, 3rd World Congress on MAAR, 2010
Congress President, 16th World Congress on IVF, 2011
Congress President, 29th Annual meeting of JSFI, 201176

●Plenary Lecture 2

Non-invasive assessment of embryo quality
Zsolt Peter Nagy
Reproductive Biology Associates
Introduction. The traditional procedure for embryo assessment is performed bymorphological evaluation which provides useful information, however, there are
numerous factors contributing to oocyte/embryo quality that are not necessarilyreflected by morphology. Review of alternative techniques on embryo viability assessment. In recent years,
there have been numerous reports on different technologies, which may aid embryo assessment by providing
additional information on the viability of the embryos. One of them is birefringence imaging which utilizes
polarized microscopy (polscope) to study the status of the meiotic spindle and the zona pelludica and canprovide information on embryo development ability. Time-lapse video monitoring of embryos is another,more complex method of visual assessment which is based on monitoring by real-time, time-lapse imaging ofindividual embryos. One of the advantages of these systems is that they minimize handling and exposure to
sub-optimal environmental effects, while enabling the continuous monitoring of embryo development andthrough analyses of developmental (cleavage) data with sophisticated software it also can provide prediction
on embryo viability. Measurement of uptake and secretion of single or specific molecules in the culture
medium has also been suggested by several studies to aid embryo assessment. Pyruvate and glucose can giveinformation on a switch from carboxylic acid to glucose metabolism that occurs in the early embryo between
pre- and post-compaction thus their concentrations can relate to the ability of the embryo to develop further.
Amino-acid measurements by ultramicrofluorometric assays or by reverse-phase high performance liquid
chromatography (HPLC) can be used to predict blastocyst formation, clinical pregnancy rate and live birth.
Soluble Human Leukocyte Antigen G (HLA G), which is believed to play a role in immuno tolerance during
pregnancy was also suggested as indicator for pregnancy outcome, as well Leptin and Platelet Activating
Factor (PAF) have also been suggested as potential markers. Oxygen consumption yet another single marker
measured by Nano and Embryo Respirometer, that can provide information relating to embryo implantation
potential. Proteomics, which refers to the study of proteins, is another technology that can be used, speciallythe method called surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI TOFMS) is particularly suited for analysis of relatively low concentrations of secreted proteins from culture media
with higher sensitivity of than other platforms, and has been described to relate embryo development to theblastocyst stage. Metabolomics is another newly introduced approach; which refers to the study of factors
present in the culture medium that are secreted by the embryo into its environment as a result of metabolomic
processes that occur within the cells. The secreted metabolites (also called secretome) are readily available
and can be collected in a simple, non-invasive way using Raman on near infrared spectroscopy. Several
studies have demonstrated strong correlation between metabolomic profile of embryo and its ability to
implant.
Conclusions. There are several novel techniques that are suggested to be able to predict embryo viability
using non-invasive approaches. Further studies will clarify, which technique(s) are best suited to aid current
embryo assessment procedures.
◆Biosketch
Dr. Nagy obtained his MD (1986) and his Ob&Gyn degrees (1996) at the Semmelweis Medical University in
Budapest. His PhD was granted at the Free University of Brussels in 1997, where he worked in the team thatdeveloped the ICSI procedure. More recently, he has contributed to the development of a highly efficient
oocyte cryopreservation procedure that gave bases to establishing the first donor cryo-bank in USA. He is
author of more than 150 publications and book chapters. Dr. Nagy serves on the board of several different
societies; as well he is Associate Editor for Human Reproduction and RBMonline. Currently, Dr. Nagy is the
Scientific and Laboratory Director at Reproductive Biology Associates, Atlanta, USA.

77

●Plenary Lecture 3

The Prospect of Embryonic Stem Cell Research
Shin Yong Moon M.D., Ph.D.
Department of Obstetrics and Gynecology,
Director of the Institute of Reproductive Medicine and Population
Medical Research Center, College of Medicine, Seoul National University
There has been no argue on the importance of stem cell research in developing of future medicine andtherapeutic bioengineering. Lots of peoples and the patients who supper from incurable diseases are
enthusiastic about clinical feasibility of stem cells since the first establishment of human embryonic stem cells(hESCs). As the strong candidate of clinical application, tissue-specific stem cells and induced pluripotent
stem cells (iPSCs), in addition to hESC, have strongly been suggested. Based on recent development of novel
cytogenetic and cytological tools, however, all of these candidate cells showed the limitations, which should
be cleared before clinical application. In the case of hESC, ESCs derived from surplus embryos is very much
likely to induce immune reaction to a recipient patient. Cellular heterogenicity in self-renewal, differentiationand localization characteristics should be controlled. Extremely low efficiency in establishing patient-specificstem cells by somatic cell nuclear transfer is critical, which concomitantly evokes serious ethical issue on its
clinical application. In iPSCs, the processes of reprogramming and adaptation to in vitro-environment cause
lots of genetic and epigenetic abnormalities including copy number variation and multiple point mutations aswell as aneuploidy. As a matter of fact, human iPSCs show more genetic abnormalities and cellular
heterogenticity than precursor fibroblasts or even hESCs and recent reports strongly concerned the acquisitionof immunogenicity to iPSC donor after reprogramming. More basically, genetic manipulation of somatic cells
for deriving iPSCs makes its clinical application impossible. Oncogene transfection to precursor somatic cells
for the cell derivation remarkably increases concerns on clinical application of iPSCs. Regardless of its
significance, inefficient isolation of tissue-specific stem cells from various adult tissues has been criticized forlong time and the establishment of homogenous stem cell population is a prerequisite factor for clinicalapplication of the stem cells. Cell niche to control stem cell fate and to mobilize tissue-specific stem cells has
not been defined yet.
In conclusion, more detailed study on genetic and cellular characteristics of stem cells will guarantee thedevelopment of stem cell engineering for clinical application, and the state-of-the-art cellular and genomic
technology can be utilized for the novel studies.
◆Biosketch
Name: Shin-Yong Moon M.D.
Born April 1, 1948
Address: Department of Obstetrics and Gynecology
Seoul National University Hospital
Yonkeun-Dong, Chongno-Gu, Seoul 110-744, Korea
Shin-Yong Moon is currently Professor of Obstetrics and Gynecology, College of Medicine, Seoul National
University Hospital, and Director of the Institute of Reproductive Medicine and Population, Medical ResearchCenter, at Seoul National University. Completing all the necessary courses and requirements for Obstetrics
and Gynecology specialists in Seoul National University Hospital, he started to study ―In Vitro Fertilization
Program‖ at Jones Institute for Reproductive Medicine, Norfolk, Virginia in United States in 1983.
After coming back to Seoul, he could have first successful IVF baby in Korea in 1985. During the last 30
years he studied Cytogenetics, Reproductive Endocrinology and Infertility and Embryonic Stem Cell. In 200178

●Plenary Lecture 3

he succeeded in establishment of ―Human Embryonic Stem Cell‖ from frozen 2 pronuclear stage
cryopreserved embryos in his laboratory.
Professor Moon has been a major contributor to the development of reproduction medicine in Korea and isactively involved in numerous local medical societies; he was President of the Korean Society of Human
Reproduction and the Korean Tissue Engineering and Regenerative Medicine Society, President of the Korean
Society for Assisted Reproduction and Emeritus President of the Korean Society of Medical Genetics.
He is also author or co-author of more than 30 papers published in leading international journals, including
Molecular Therapy, Stem Cells, Human Reproduction and Fertility and Sterility.‖79

●Plenary Lecture 4

Preimplantation genetic diagnosis: an update
Alan H. Handyside
London Bridge Fertility, Gynaecology and Genetics Centre
Twenty one years on from the first pregnancies and births in July, 1990
preimplantation genetic diagnosis (PGD) is now clinically well established as analternative to prenatal diagnosis. As genome wide analysis becomes quicker,
cheaper and more widely available over the next decade, genetic testing of both patients and their embryos
will be revolutionised. Ultimately it may even be possible to sequence the entire genome of an embryo though
it is more likely that targeted methods will be more cost effective. The benefits to patients are that this will
enable accurate testing of embryos not only for virtually any chromosomal or single gene defect identified in
the parents but also for genetic defects formed spontaneously in the gametes and early embryo, many of which
are likely to compromise viability and development. As the unveiling of the first personal genomes has
demonstrated, however, genome wide analysis raises challenging clinical, ethical and social issues.
Increasingly it may be possible, for example, to provide information about physical characteristics andpredisposition to common diseases and late onset disorders. On the other hand, thousands of genetic variants
are being discovered for which the consequences are unknown. There is a danger that this will increasedemands for PGD from prospective parents who want to know (or control) how their child will turn out andwhether they will be healthy. Future regulation of PGD needs therefore to focus on not only what inheritedconditions should be allowed for embryo selection but also what genetic information should be provided topatients about those embryos that are selected for transfer.
◆Biosketch
Background in mouse and human embryology and genetics with over 200 publications. Created one of thefirst mouse transgenic mice using embryonic stem cells. With Prof Lord Robert Winston, achieved the firstpregnancies following IVF and PGD worldwide in 1990. First Chairman ESHRE Special Interest Group in
Reproductive Genetics and founder and first Chairman of the ESHRE PGD Consortium.

80

●Plenary Lecture 5

Preservation of Fertility
Sherman J. Silber1, Noriko Kagawa2, Masashige Kuwayama2
1
Infertility Center of St. Louis, 2Kato Ladies Clinic

Objective: Cryopreservation of ovarian tissue is widely performed to preserve
fertility in cancer patients, but its efficacy has been difficult to assess because
most patients do not request transplantation of the thawed tissue for over a decade. Our objective was toevaluate the long-term results of transplantation of fresh ovarian versus frozen ovarian tissue in patients withovarian failure (OF), to evaluate its usefulness in preservation of fertility for cancer patients.
Design: Seven year follow-up of a consecutive series at one center of women with OF who underwent either
fresh (11) or frozen (4) ovary transplantation, and evaluation of the tissue of 62 patients who underwent
ovarian cryopreservation, but not yet transplantation.
Materials and Methods: 12 women with ovarian failure were recipients of 15 ovarian tissue transplants, 11fresh from a sister, and 3 frozen autotransplants. In 9 of the fresh cases, the sisters were identical twins, and in
2 cases, they were non-identical siblings. In 4 women, the cause of OF was cancer chemotherapy andradiation. In the other 8 women, the cause was idiopathic premature OF. A small portion of tissue was tested
for post-thaw viability in all cases of ovarian tissue freeze.
Results: There have been 17 pregnancies, and 14 healthy babies (11 from fresh transplant and 3 from frozen
tissue), with 3 miscarriages. 5 babies were delivered to women who had received pelvic irradiation. With freshtransplants, menstruation resumed from 63 to 130 days post-transplant, whereas with slow frozen transplants,
menstruation resumed after 147 days. Fresh transplants continued to function from 2 years to over 7 years (4of the 8 cases) depending on the donor's pre-operative antral follicle count. The average duration of function
was over 5 years. Frozen (slow freeze) grafts, however, functioned for 1-1⁄2 to 2 years. Examination of
vitrified grafts revealed no difference between pre-freeze and post-freeze viability.
Conclusions: Ovarian cortical grafting, fresh or frozen, particularly with vitrification gives robust results withhealthy babies, and good, long-term function.
◆Biosketch
Dr. Sherman Silber is one of the world's leading authorities on IVF, microsurgery, vasectomy and tubal
reversal, egg and embryo freezing, ovary and testis transplantation, and the reproductive biological clock. He
is director of the Infertility Center of St. Louis at St. Luke's Hospital in St. Louis, and author of thebest-selling How To Get Pregnant.

81

●Plenary Lecture 6

The role of reproductive surgery in the era of IVF & ART
Victor Gomel
Department of Obstetrics and Gynecology,
Faculty of Medicine, University of British Columbia,
Vancouver BC. Canada
The indications for reconstructive surgery in the era of IVF and ART are two-fold: 1.
Primary treatment of tubo-ovarian disease, adhesions and other pelvic pathology to give the patient theopportunity to achieve spontaneous conception; and 2. Treatment of uterine and other conditions that would
adversely affect the outcome of assisted reproduction
Tubo-ovarian disease, adhesions and other pelvic pathology are the principal cause of infertility inapproximately one third of cases. While, reconstructive surgery was at one time the only treatment option, thisis no longer the case. Improvement in the outcomes, simplification of the techniques and much wider
availability of the services of in vitro fertilization (IVF) and assisted reproduction (ART), provide suchcouples with a realistic therapeutic alternative.
Further to significant improvement in IVF results in the nineties, the pregnancy rate per cycle initiated appears
to have stabilized in the USA at around 28% during the last several years (2002-2008). Success with transfer
of cryopreserved embryos has also improved significantly. The introduction of intra-cytoplasmic sperm
injection (ICSI) proved to be a panacea in the treatment of male infertility. Indeed the outcomes of IVF withICSI produce similar outcomes in couples with and without male factor.
This enormous progress in the live birth rates with ART has been accompanied with the commercialization of
the technology and its services, all over the world. In parallel fashion there has been a significant decline in
the practice and teaching of reproductive surgery. IVF now is offered as primary treatment option, in mostcases of tubal factor infertility. These changes have occurred despite the major progress that gynecologic
surgery has experienced; introduction of microsurgical techniques and the predominant use of laparoscopic
surgical access that readily permits the application of these techniques in reproductive surgery.
ART is associated with a tremendous increase in preterm births and its important sequelae of perinatal
mortality and morbidity, including cerebral palsy. The rates of preterm births for singletons, singletons frommultiple fetuses, twins, triplets and greater order of multiples are 12.5%, 18.3%, 63.3% and 95.3%
respectively. The perinatal mortality was 1.9% in the in-vitro fertilization group almost double that of thecontrols. In the USA, in 2007, of the 36,079 pregnancies resulting from ART 28.8% were twins and 3.7%
were triplets and greater order of multiples; and of the resulting 29,556 births 29.4% were twins; greater orderof multiples decreased to 1.8% either by fetal reduction and/or spontaneously. The large proportion ofmultiple births, with the associated increased obstetrical complications, neonatal complications and deaths,
causes great societal costs and significant financial burden and emotional costs for parents.
Based on the current US outcomes of a birth rate of 28% per initiated cycle, the cumulative probability of live
birth after 3 cycles of treatment would be around 52%. However, several studies have shown conclusively thata large percentage of couples do not wish to complete 3 cycles of IVF, even when these are paid by the stateor covered by insurance. Reports from France, where IVF is covered by the state, corroborate with thesefindings. Others refuse to have IVF for religious or ethical reasons, and many would find the cost of IVF
prohibitive, since the procedure is not covered in many states and countries, while other treatment modalities
are, as is the case in British Columbia in Canada.
For the infertile woman with tubo-ovarian and pelvic pathology there are only two realistic options to achievea pregnancy: reconstructive surgery or IVF. The presence of a credible alternative, in IVF, permits the
reproductive surgeon to operate on cases with a better prognosis, which was not the case before the end of the82

●Plenary Lecture 6

eighties. We have known for a long time that one of the important factors influencing surgical outcome was
the degree of tubal damage and extent of pelvic disease and adhesions. Operating on patients with better
prognosis translates in superior outcomes, which has been well demonstrated.
With appropriate preparation, many reconstructive procedures can be performed during the initial diagnostic
laparoscopy using microsurgical techniques: salpingo-ovariolysis, salpingostomy and even tubo-tubal
anastomosis for reversal of sterilization. This is also applicable to other pelvic pathology, such as
endometriosis, uterine myoma etc. Intrauterine conditions may be treated performing a concomitant
hysteroscopy. In the absence of any other infertility factors, laparoscopic salpingo-ovariolysis yields
intrauterine pregnancy rates of 50% to 60%; and salpingostomy yiels rates ranging from less than 20% to 50%,
depending largely on the degree of tubal damage and the extent and severity of adhesions.
Microsurgery is ideal for tubo-tubal anastomosis and produces excellent results that are principally dependent
on the length and status of the reconstructed tube. In cases of reversal of sterilization, live birth rates of 60%
to 80% can be achieved. In cases of true pathologic proximal tubal occlusion microsurgical tubo-cornual
anastomosis yields an intrauterine pregnancy rate of about 50%. Microsurgery also permits reconstruction incomplex situations, for example a tubo-ovarian transposition, with preservation of their vascular pedicles. In
such cases, the prognosis that the surgical reconstruction offers is not necessarily proportional to the technicaldifficulty of the procedure.
Assisted reproductive techniques are being used increasingly as primary treatment for infertility. This islargely the result of the commercialization of the ART technology and its services. The industry is wellcapitalized and is lucrative. Yet it is important to stress that even if couples were to undergo three successive
cycles of IVF, nearly 50% would fail to obtain a baby.
Many patients need to have reproductive surgery before IVF for various conditions such as myomas, adnexal
tumors, endometriosis, etc. Yet, there has been a significant decline in the practice and teaching ofreconstructive surgery and microsurgery in gynecology. Such teaching and practice made the gynecologist amore refined surgeon, attributes that would be regrettable to loose.
As evident from the preceding, reconstructive surgery when well performed, in properly selected cases offers
satisfactory results. Furthermore, it offers the couple the opportunity to attempt a pregnancy over a long
period of time and to conceive more than once out of the same procedure. This and the available data suggestthat there should be a real place for reconstructive surgery. The preservation of this place will require aconcerted effort on the part of the teaching institutions.
The development of operative laparoscopy, tubal microsurgery, and IVF in the last 30 years has significantly
improved the outlook of couples suffering from tubal infertility. These are complementary approaches that canbe used singly or in combination. When both alternatives are equally available to the patient and used a muchgreater overall cumulative birth rate can be obtained.
In the preface of my book, Microsurgery in Female Infertility, published by Little Brown, in early 1983, I
wrote: ―This manuscript has been completed during a time of rapid change and expansion with theunderstanding that it represents not an endpoint but merely an accounting at a given point in time. Further
developments are also occurring in the area of IVF and embryo transfer (IVF & ET), which will undoubtedly
produce improved results. Nonetheless, I do not consider the techniques of microsurgery on the one hand and
IVF & ET on the other as competitive; on the contrary, I see them as complementary, enabling us to achieve agreater success rate among those patients presenting with complex fertility problems.‖ This statement is stillvalid today.
◆Biosketch(For Prof. Gomel‘s Biosketch please refer to his abstract for Concurrent Symposium C-4.4.)83

●Plenary Lecture 7

Recent advance of controlled ovarian stimulation
Klaus Diedrich
Department of Obstetrics and Gynecology of the University of Lübeck,Germany
Ovarian stimulation contributes to the overall effectiveness of in vitro fertilization
treatment. However, ovarian stimulation is also associated with health risks, adverse events, treatment burden
for the patient and high financial costs. Ovarian stimulation therefore needs to be continuously improved. In
this literature review, three important new developments in the field of ovarian stimulation have been selected
for discussion. Human chorionic gonadotropin as the triggering agent for ovarian hyperstimulation syndrome(OHSS) can now safely be replaced with a bolus dose of a gonadotropin-releasing hormone agonist. This has
been shown to reliably prevent OHSS, the most serious complication of ovarian stimulation. To reduce theinjection frequency of gonadotropins, a long-acting follicle-stimulating hormone molecule (C-terminal peptide,
FSH-CTP) has been developed and tested in a large set of clinical trials. It was shown that long-acting
FSH-CTP is able to stimulate the ovaries for 7 days at doses of 150 and 100 µg, respectively, and that theoutcome in terms of pregnancy likelihood is similar to conventional gonadotropin stimulation by daily
injection. Orally active non-peptide mimetics of luteinizing hormone and FSH are currently being developed.
However, no data on the administration to humans have been published to date, and only scarce data on invitro and animal experiments are available.
◆Biosketch
Prof. Dr. med. Dr. h.c. mult. Klaus Hermann Rolf Diedrich
Birth:
28.04.1946
Birth-place: Nordenham
Parents:
Dr. med. Hermann Diedrich, M. D.
Rita Diedrich (maiden-name: Ehrlich), lady of the house
Education:
1966:
High school graduate at the Matthias-Claudius-Gymnasium in Hamburg
1966:
Start of studies in human medicine at the University of Hamburg
1967:
Naturwissenschaftliche Vorprüfung in Hamburg1968:Ärztliche Vorprüfung
1972-73:
Junior house officer in surgery and internal diseases
1973-74:
Military service as M. D.
1974-78:
Senior house officer in OB/GYN at the university clinic of Hamburg
1979:
Consultant at the department of OB/GYN at the university clinic of Lübeck
1981:
Ph. D. in Obstetrics and Gynecology
1982:
Marriage with Christa Diedrich, M. D.
1983:
Birth of a son
1984:
First consultant at the department of OB/GYN of the university clinic of Bonn;
University Professor
1984:
Founding member of the European Society of Human Reproduction and Embryology(ESHRE)
1985-91:
Secretary of the European Society of Human Reproduction and Embryology (ESHRE)

84

●Plenary Lecture 7

1991-93:
1993:
1993:
1994-2000:
2001:
2000:
2002:
2004:
2006:
2006:
2006:
2009:
2009:
2009:
2010:

Chairman-elect of the European Society of Human Reproduction and Embryology(ESHRE)
Head of department of Obstetrics and Gynecology of the University Clinic of Lübeck
President of the European Society of Human Reproduction and Embryology (ESHRE)
Secretary of the German Society of Obstetrics and Gynecology (DGGG)
Member of the German Academy of Science Leopoldina
Vice President (chairman-elect) of the German Society of Obstetrics and Gynecology(DGGG)
President of the German Society of Obstetrics and Gynecology (DGGG)
2. Vice President of the German Society of Obstetrics and Gynecology (DGGG)
Vice Dean of the University of Lübeck
Fellow of the Royal College of Obstetrics and Gynecology
Foundation of the German-Greek Society of Obstetrics and Gynecology by Prof. Agorastos(President) and Prof. Diedrich (Vizepresident)
Medical Director of the University Hospital of Lübeck
Doctor honoris causa of the Aristoteles University of Thessaloniki
Doctor honoris causa of the University of Alexandropoulis
Medical director of the University Hospital Schleswig- Holstein, Campus Lübeck

Member of 18 national and international Editorial boards.
Publications: 480 in national and international journals.

85

●Plenary Lecture 8

Tending to Male Factor Couples
Gianpiero D. Palermo, M.D., Ph.D.
The Ronald O. Perelman & Claudia Cohen Center for Reproductive
Medicine, Weill Cornell Medical College, New York, NY, USA
In the USA infertility affects about 13.3%, approximately 7.3 million, of all
couples of reproductive age, but only about 1.5 million of those benefit fromassisted reproductive technologies (ART). Unexplained and combined male and female factor infertility
accounts for 34%, the remainder being evenly attributed to female (33%) or male (33%) problems.
Since the inception of in vitro fertilization (IVF) in 1978, it has become evident that half of the couples failed
to achieve fertilization because of sperm-related problems. This stimulated the development of several
micromanipulation procedures that resulted in the establishment of intracytoplasmic sperm injection (ICSI), a
reliable procedure through which to achieve fertilization. ICSI has now been employed successfully even incases where spermatozoa can only be aspirated from the epididymis and/or the testis of azoospermic men.
The causes of male infertility include the presence of a varicocele in some 30% of such men, followed by
unexplained reasons in 25%, and accessory gland abnormalities and cryptorchisdism in 13% and 9%,
respectively. The obvious problem of azoospermia due to vas obstruction or testicular failure is seen in about
9% and 14% of male factor couples, respectively.
As a further element, chromosome anomalies are a significant cause of infertility, pregnancy loss and the birthof children with mental and physical disorders. Understanding the genesis of such anomalies requires the
direct study of human spermatozoa as such. Since the great majority of errors occur during meiosis, analysis atother spermatogenic stages will minimize the ascertainment bias due to the loss of chromosomally abnormal
conceptuses during development. Even males with normal peripheral karyotype may have chromosomal
abnormalities in their germ cells, these often arising through instances of nondisjunction during
spermatogenesis. Therefore, it is important to screen for sperm aneuploidy.
The root of poor sperm production is often genetic. Male subfertility has been associated with a higher
incidence of genomic defects, ranging from aneuploidy to Yq microdeletions, and concerns have been raised
as to the risk of transmitting these to the offspring. For this reason, screening for such defects can be an
important adjunct to appropriate counseling prior to ICSI treatment. So far, only a few reports of father/son
cohorts have evaluated the heritability of mutations associated with male factor infertility, particularly thewell-being of the children. Although early studies on neonatal outcomes failed to reveal any differences
between ICSI and IVF babies, some recent reports have claimed that there is a greater incidence ofabnormalities in ART children, and also an increased risk of low birth weight (and very low birth weight)
when compared to those conceived normally. Anxieties have also been voiced in regard to rare imprinting
disorders, as well as cancer following ART.
Since 1993, ICSI has been employed in over 30,000 ART cycles at our center and has resulted in positive
pregnancies in 40.1% of cases, based on the presence of at least one fetal heartbeat, comparable to thoseobtained with conventional IVF.
A total of 902 cycles were performed with epididymal spermatozoa and 1,143 cycles with testicularspermatozoa. Both fertilization (71.5%) and clinical pregnancy rates (52.5 %) were higher (P < 0.001) withepididymal versus testicular samples (TESE fertilization 56.9% and clinical pregnancy rate 40.8%). When we
assessed whether or not cryopreservation affected clinical outcome and observed that fresh MESA (61.9%)
had a higher clinical pregnancy than TESE (41.3%; P = 0.04) and a similar result was observed when
specimens were cryopreserved (frozen MESA 47.6 versus frozen TESE 36.1%; P = 0.003). When we lookedwithin a particular surgical retrieval method, the clinical pregnancies with TESE was comparable regardless of86

●Plenary Lecture 8

the sample being fresh or cryopreserved. However, fresh MESA showed a higher pregnancy rate incomparison to its frozen counterpart (61.9 vs 47.6%, P = 0.0001).
In 8,293 patients a viable fetal heart was observed by ultrasound, where 898 patients miscarried, aborted, or
had an ectopic. The ongoing pregnancy rate was 36.1% per retrieval (7,444/19,226) and 38.7% per
replacement procedure (7,444/10,676). So far, we have 8,695 neonates born from 6,537 deliveries.
In reproductive medicine, ART has played a major role in the treatment of couples with all forms of infertility,
and in this arena ICSI has distinguished itself in shedding light on the intricate mechanisms of gamete
interaction, fusion, and early embryo development. In addition, ICSI has been a research tool through whichto study the male gamete not only in regard to its role as the carrier of the male genome but also in its ability
to trigger ooplasmic calcium release and its provision of the precursor of the mitotic spindle — of paramountimportance for the generation of a healthy embryo.
The recent trends in ART have also raised concerns regarding genetic and epigenetic risks to the offspring.
This experience has enabled us to better counsel couples contemplating reproductive treatment, tometiculously monitor the safety of all procedures applied, and, not least, to carefully assess the physical andpsychological wellbeing of the children involved.
◆Biosketch
Gianpiero D. Palermo, M.D., Ph.D.
Gianpiero D. Palermo is the developer of intracytoplasmic sperm injection (ICSI), the revolutionary procedurethat has been able to virtually cure male infertility. He established the ICSI programs with Prof. André Van
Steirteghem at the Brussels Free University in Belgium and later with Prof. Zev Rosenwaks at Cornell
University in New York. There are over 2 million babies born from ICSI worldwide.
Dr. Palermo completed his clinical training in Obstetrics and Gynecology at the University of Bari in Italy,
attended the Masters and Ph.D. programmes in Brussels Free University and is currently completing
additional post-doctoral training in New York and Melbourne. Dr. Palermo has won many prestigious prizes
and awards for his pioneer work in Reproductive Biology, including two presentation awards from Serono
Laboratories, the Barbara Eck Menning from RESOLVE, the Shackman Memorial Lecture at Johns Hopkins,
the Buckeye Lecture of the ASA, Jacob Heskel Gabbay Award in Biotechnology and Medicine, and The
Crystal Tube Award.
Dr. Palermo has delivered almost 300 lectures before international audiences, presented over 250 abstracts,
several book chapters, and authored/co-authored over 100 peer-reviewed articles. Since 1993, he has been the
Director of the ICSI Program at The Ronald O. Perelman & Claudia Cohen Center for Reproductive Medicine
and Professor at the Weill Cornell Medical College. He leads a team of talented researchers actively involved
in molecular and genetic aspects of fertilization, follow-up of ICSI babies, genetic aspects of male infertility
as well as devising new procedures to treat age-related female infertility, harvesting and differentiation ofembryonic stem cells, and in vitro maturation of male germ cells.

87

●Plenary Lecture 9

In-Vitro Maturation (IVM) for fertility treatment and preservation offertility
Seang Lin Tan
McGill University
IVM is a relatively new technology with limited clinical experience compared toconventional IVF. IVM should be regarded as a complementary assisted reproductive technology of IVF thatprovides unique opportunities for patients at high risk of OHSS, those with unexpectedly hyper- or poor
responses during controlled ovarian hyperstimulation, those with recurrent unexplained IVF failures, andthose who are facing imminent gonadotoxic chemotherapy. Essentially, all ART laboratory procedures can be
performed with in-vitro matured oocytes if the need arises. The first successful IVM cycles combined withpreimplantation genetic screening and percutaneous testicular sperm aspiration have already been reported byour team. We have found aneuploidy rates to be similar in IVM and IVF embryos. IVM has enabled
successful treatment of patients with empty follicle syndrome in previous stimulated IVF cycles. Patients canundergo several IVM cycles and we previously reported a series of patients who achieved repeated live births
with IVM treatment. Currently, IVM represents the least invasive and the simplest option, and with further
improvement in success rates, could become the ultimate patient-friendly protocol. Current clinical pregnancy
rates per cycle started reach 50% in young women. Improved treatment methods have resulted in a steadyincrease in cancer survival rates over the last decades, and a growing number of women are facing the risk ofinfertility resulting from gonadotoxic oncologic treatment. Patients exposed to gonadotoxic agents for thetreatment of non-oncologic diseases such as systemic lupus erythematosus, who are undergoing surgery forendometriosis and who suffer from genetic disorders such as Turner syndrome and fragile-X pre-mutation
face similar risks. With an increased awareness of the options available, more women are being offered andare utilizing fertility preservation technologies. The methods can differ for women with medical conditions
and for those who wish to defer child bearing for social reasons in the absence of a medical condition. IVF
followed by embryo cryopreservation is regarded as the only established method for fertility preservation infemale cancer patients; however, concerns about IVF/embryo freezing include: (1) possible delay in treatmentof the primary disease due to ovarian stimulation in relation to onset of the next menstrual cycle (2) exposure
to supraphysiologic estrogen levels induced by ovarian stimulation (3) the need for a male partner or donor
sperm for embryo production (4) legal, ethical, religious issues related to embryo cryopreservation in
general.IVM avoids treatment delay or exposure to increased estradiol levels associated with IVF and,combined with embryo or oocyte vitrification, provides previously unavailable options, such as immature
oocyte collection in the luteal phase for some patients, and improves the services provided by a fertility
preservation program. Primary-care physicians and oncologists should be made aware of the available fertility
preservation options in order to allow referral of their patients, if desired, to an ART center that offers the fullrange of fertility preservation options. We have preserved fertility for over 200 women with various medical
conditions. In a clinical trial of IVM and oocyte vitrification, we achieved a live-birth rate of 20% and thebirth of the first four healthy babies.
◆Biosketch
Dr. Tan is Professor and James Edmund Dodds Chair in ObGyn at McGill University, internationallyrenowned infertility expert and a pioneer of IVM and oocyte vitrification. He is founding director of McGill
Reproductive Center, founding President of GCARM and ISIVM, founding Treasurer of ISIVF and Medical
Director of the Montreal Reproductive Centre. He led the team that produced the first air transport IVF/ICSI
pregnancies in the world, first IVM twin birth, first IVM/PGD birth, first 4 IVM/oocyte vitrification (OV)88

●Plenary Lecture 9births and first serial OV and PGD birth. His many awards include the Howard Eddey, MRCOG, Benjamin
Henry Sheares and RCOG Singapore Lecture Gold Medals, Resolve Award, John Collins Lectureship,
CFAS/ESHRE Exchange Speaker Award, Montreal Arts and Trade Medal and the College of ObGyn Gold
Medallion lectureship. He has published 12 books, 248 original articles in many major journals, including
Nature, The Lancet and NEJM, and 92 book chapters and reviews.

89

CONCURRENT SYMPOSIA

90

●Concurrent Symposium C-1.1

Androgenic spectrum concept of PCO morphogenesis
Takahide Mori
Academia for Repro-regenerative Medicine,
Daigo Watanabe Clinic, Kyoto, Japan
Polycystic ovarian (PCO) morphology is commonly observed both in women withpolycystic ovary syndrome and with regular menstruation. Hormonal regulation for morphogenesis has not yet
been scrutinized. In 1991, we reported that PCO syndrome (PCOS) could be divided into three discrete types
based on pituitary-ovarian androgenicity; classical Stein-Leventhal syndrome, hyper-LH with hyperandrogemia and simple hyper-LH. The world-widely accepted classification of Rotterdam consensus on
diagnostic criteria seems ignored androgen-producing potency by theca cells under LH control. Therefore, we
examined In recognition of complementary interaction of androgen-induced expression of FSH receptor on
granulose cells with FSH-augmented expression of the androgen receptor on granulosa cells in early antral
follicles until selection of the dominant follicle, a possible correlation of antral follicle count (AFC) andpituitary-ovarian androgenic function was investigated in 180 infertile women during 3-5 days of themenstrual cycle under the understanding that PCO morphology is composed of a mixture of healthy andatretic antral follicles. AFC, taken as an indicator of PCO morphogenesis, was compared with clinical signs ofandrogen excess and laboratory measures of baseline LH, LH/FSH ratio, and total testosterone concentrations
as parameters for pituitary-ovarian androgenic function. Six discrete types with decreasing pituitary-ovarian
androgenic activity were identified in parallel to the declining number of AFC: Type I (classicalStein-Leventhal syndrome), Type II (hyperandrogenism), Type III (singular hyper-LH), Type IV (cryptic
hyperandrogenism), Type V (relative LH dominancy) and Type VI (relative FSH dominancy). Since AFC
reflects the basal number of antral follicles generated, these six different types with each characteristic
androgenic pattern of the pituitary ovarian androgenic activity is thought to represent dynamic wave of PCO
morphogenesis under the regulation of enhancing androgenic activity of the pituitary-ovarian axis.
Considering that antral follicles over 3 to 5 days of the menstrual cycle consist of newly emerged selectable
healthy follicles plus atretic antral follicles that are left unovulated from previous cycles, we proposed the
hypothesis that the six identified types of PCO with increasing androgenic function represent thefolliculogenetic spectra along which PCO morphogenesis may proceeds in normally cycling as well as PCOS
patients, though additional factor(s) may be involved in the latter.
◆Biosketch
Education: MD degree from Kyoto Univ in '60, PhD degree in '68.
Research carrier: Follicular steroidogenesis in Miami, and reproductive immunology at Population Council.
Academic carrier: Chairman of OB/GYN Dept Univ of Tokushima in '81. He organized WHO Workshop on
Reference Bank Sera during the term.
Academic interest: Since the first discovery of progesterone suppression on lymphocytes in '77,
endocrine-immune interaction in reproduction became his major academic interest with remarkable success.
Academic achievements: A total of 1310 articles including 516 English papers.
IVF: Establishing the first IRB at Univ of Tokushima on IVF into Japan which was highly evaluated as amonumental attempt in the country. He organized 8th World Congress on IVF as President in Kyoto in '93.
Post-retirement activities: After retirement from Kyoto Univ. he continues research and practice at
Daigo-Watanabe Clinic, concurrently serving as Director of Academia for Repro-regenerative Medicine.

91

●Concurrent Symposium C-1.2

Do phenotypic differences in PCOS affect fertility treatment andoutcomes?
Roger A. Lobo M.D.
Professor Obstetrics and Gynecology
Columbia University, New York, NY, USA
President American Society for Reproductive Medicine
The diagnosis of PCOS has broadened considerably in recent years, with many different phenotypes now
being possible, including that of ―ovulatory‖ women. Phenotypic differences vary among regions of the world,with the more severe (or classic) phenotype more commonly seen in the US, and also being the focus of
most studies on PCOS. If hyperandrogenism is considered to be a key feature for the diagnosis, then there isevidence that this feature may have a negative effect on fertility even in ovulatory women. This may be
manifest in subtle abnormalities in ovarian function as well as in endometrial maturation. Phenotypic variation
also may influence the miscarriage rates in PCOS, which have been considered to be increased, yet may only
be increased in a subset of the disorder, namely obese women with a more severe phenotype. The
Thessaloniki ESHRE/ASRM guidelines for treatment of infertility in PCOS stated that clomiphene is still firstline therapy, followed by either ovarian ―drilling‖ or gonadotropin therapy, and then followed by IVF-ET as
third line therapy. These guidelines discouraged the use of metformin except for glucose intolerance and theuse of aromatase inhibitors. However, depending on the phenotype involved, there is ample accumulating
evidence that metformin and letrozole both have a role in treating women with PCOS. There remains
controversy as to whether oocytes from women with PCOS are normal or abnormal, although it has been
established that IVF pregnancy rates are comparable to that of women with tubal infertility. Here, it appears
that phenotypic variability does not affect IVF outcomes except for the influence of increased BMI/obesity
being negative. Uses of metformin and antagonist therapy appear to have a role in reducing the risk of OHSS
in PCOS. Poorer obstetrical outcomes and a higher neonatal death rate have been reported in women with
PCOS, but this is probably related to the more severe phenotypes of PCOS, and is less evident in ovulatory or
non hyperandrogenic women.
◆Biosketch
Rogerio A. Lobo, M.D. received his medical degree from Georgetown University Medical School in
Washington, D.C., and completed his residency in Obstetrics and Gynecology at the Chicago Lying-in
Hospital at the University of Chicago. He went on to complete a clinical research fellowship in the Division of
Reproductive Endocrinology and Infertility at the University of Southern California Medical Center in Los
Angeles.
He has since held numerous teaching positions, including Assistant and Associate Professor in the Departmentof Obstetrics and Gynecology at the University of Southern California in Los Angeles, where he was later
promoted to full Professor. In 1995 he came to the Columbia University College of Physicians and Surgeonsin New York City where he was named the Willard C. Rappleye Professor of Obstetrics and Gynecology, andChairman of the Department. He was also appointed Director of the Center for Reproductive Sciences at theCollege of Physicians and Surgeons and Director of the Sloane Hospital for Women, Columbia University
Medical Center, one of the most prestigious women‘s health centers in the country.
In addition to a successful career in academics, Dr. Lobo has excelled in other aspects of professional
medicine, serving as the Director of the Reproductive Endocrinology and Infertility Training Program at the92

●Concurrent Symposium C-1.2

University of Southern California for 11 years. He has provided outside consulting services for many large
pharmaceutical laboratories. He has functioned as Editor, Editorial Board member and/or Consultant for over
30 peer-reviewed medical journals in his field, and has authored over 400 articles and 20 books. Dr. Lobo
helped found the Journal of the Society for Gynecologic Investigation and served as Editor-in-Chief from its
inception until July, 2006. He was President of the Society for the years 1997-1998. He has contributed
Chapters to and edited several important medical textbooks. His book Treatment of the Post-Menopausal
Women is in its Third Edition and published this year. (2007) Another text, Menopause, was published inearly 2000.
Dr. Lobo has done extensive research in various areas of reproductive endocrinology and infertility. His
primary research interests are in reproductive endocrinology, specifically in hyperandrogenic disorders andpolycystic ovary syndrome. He also carried out extensive research in gamete biology, induction of ovulation,
IVF, and estrogen metabolism and the treatment of postmenopausal women. He has excelled in his areas ofclinical interest and practice which include reproductive endocrinology, infertility, and menopause. In 1910 hebecame President of the American Society For Reproductive Medicine.

93

●Concurrent Symposium C-1.3

Laser assisted ICSI for oocytes matured in vitro from PCO patients
Muchsin Jaffar, Yuslam. E Fidianto, Hadi Sjarbaini,
Soegiharto Soebiyanto, Dianing Amalia, Malvin Emeraldi
Family Fertility Center, Family Children and Maternity Hospital, Jakarta,
Indonesia
INTRODUCTION: ICSI is a microinvasive procedure that is often used in In Vitro Maturation (IVM)
treatment to handle zona changes of in vitro matured oocytes that have resulted from long term culture. In
ICSI procedure, penetration of the injection pipette into the oocytes can sometime very difficult when there isresistance of the zona pellucida and/or the fragility of the oolemma. Application of high pressure in thissituation can damage the oocytes. The high degeneration rate may result in losing the cycle, especially whenvery few oocytes are available because there are no embryos available for transfer. In order to avoid suchdeformations and to minimize degeneration of the oocytes, in this study spermatozoa was injected into the
oocyte through a microhole on the zona pellucida drilled by laser beam just before ICSI to facilitate
penetration of the injection needle into ooplasm.
MATERIAL & METHODS: As many as 389 immature oocytes were collected from 32 PCO patients (36cycles) from November 2008 to December 2010. All female patients received pretreatment with FSH andhCG priming. The oocytes were cultured in maturation medium for 24 to 48 hours in the 5% CO2 incubator.
After the oocytes reached metaphase II, they were grouped according to their morphology at the insemination
time into the study group [oocytes with less favourable morphology where laser-assisted (L-ICSI) was used as
method of insemination] and the control group [oocytes with more favourable morphology whereconventional ICSI (C-ICSI) was used].
RESULTS: A total of 315 MII oocytes were obtained after in vitro maturation for 48 hours. Of this, 299
oocytes were subjected to ICSI [L-ICSI (n=224); C-ICSI (n=75)]. Oocytes treated by L-ICSI survived better[95.1% (213/224) versus 85.3% (64/75), P=0.005] and tended to give higher fertilization rate [61.2%(137/224) versus 50.7% (38/75), P=0.071] but lower number of good embryos [2-4 cells on the second day ofdevelopment, fragment <10%; 62.8% (86/137) versus 68.4% (26/38), P=0.179] compared to oocytes treatedby conventional ICSI. Four healthy babies were born from 9 pregnancies achieved after transferring embryosfrom L-ICSI, C-ICSI or mixed.
CONCLUSIONS: Laser asissted ICSI by drilling microhole on the zona pellucida just before ICSI is a safe,
simple and easy to perform to facilitate penetration of the injection needle into ooplasm. Our preliminary
results show that laser assisted ICSI could give higher oocyte survival rate in patients who have higher risk for
oocyte degeneration such as in IVM .
◆Biosketch
Dr Muchsin Jaffar is the Chief Embryologist and Director of Family Fertility Center, Jakarta Indonesia. He
has been trained at some prominent IVF centers in Europe such as: Academisch Ziekenhuis Dijkzigt,
Rotterdam, The Netherlands in 1987; Academisch Ziekenhuis-Vrije Universiteit Brussel, Belgium in 1992
and at National University Hospital, Singapore in 1994. He is one of the pioneer of IVF in Indonesia with over
20 years experience in the area of Infertility and IVF practice. He is involved in the developments of many
IVF centers in Indonesia. His work has been significant for the advancement and growth of the field ofAssisted`Reproductive Technology in Indonesia. Dr Muchsin Jaffar is a founder member and has served on anumber of senior comittees of the Indonesian Society of In Vitro Fertilization. His current interests are in IVF,
IVM, ICSI, PGD as well as in Male Factor Infertility.

94

●Concurrent Symposium C-1.4

The use of GnRH antagonist in the controlled ovarian stimulation for
IVF in PCOS patients
Khaled Mahmoud
Centre of Reproductive Medicine and Prenatal Diagnosis, Tunis-Tunisia
Polycystic Ovarian Syndrome (PCOS) is a common endocrinopathy that affects 5
to 10% of Women of reproductive age. Regarding diagnosis, a consensus wasreached on the criteria that identify and define the syndrome (The Rotterdam PCOS consensus workshop
sponsored ESHRE/ASRM- group-2004), but as regards the treatment of infertility caused by this syndrome,the debate remains wide open despite the 2008 publication of the work of Thessaloniki ESHRE /ASRM-sponsored PCOS Consensus Workshop Group.
Ovarian stimulation of PCOS patients entering in IVF program is difficult to control because we should take
into account not only of the quantitative aspect with a production of an excessive number of oocytes and the
major risk of OHSS but also of the qualitative aspect to having a good quality of oocytes that would give goodembryos.
It was reported that, compared with GnRH agonists, GnRH antagonists that have been introduced recently inthe controlled ovarian stimulation, offer several advantages that could secure these women with PCOS
specially as in term of live birth rate the 2 products are identical (kolibiannakis, 2006) The main advantages
are: (i) Requirements for exogenous gonadotropins are reduced, rendering ovarian stimulation less costly. (ii)
Duration of ovarian stimulation protocols is shortened, improving patient discomfort. (iii) The incidence ofOHSS is lower.
In our center we conducted a randomized controlled trial from January 2004 to June 2009 included 420 PCOS
patients who came for treatment with IVF / ICSI. All patients received oral contraceptive pill (Diane35)
starting on day 2 or 3 of the cycle prior to ovarian stimulated cycle for IVF/ICSI either by a long GnRH
agonist down regulation protocol (n=210, agonist group) or by a flexible GnRH antagonist protocol (n=210,
antagonist group).
The starting dose of rFSH (GonalF) was 150IU per day for all patients in both group, this dose was adjusted
after 4 or 5 days of stimulation depending on the ovarian response.
The triggering of final maturation of oocytes was made according to usual criteria by rHCG (Ovitrelle 6500
IU) or by 0.2 mg of Decapeptyl in case of hyper-response with E2 greater than 3500 pg / ml predicting womenat risk for ovarian hyper stimulation syndrome.
Both groups of patients had similar baseline characteristics regarding their age, hormonal profile and durationof infertility.
The analysis of our results found that the COS with rFSH and antagonist in PCOS patients is as efficacy asrFSH and agonist and also more safe: less duration of stimulation, less consumption of gonadotrophins andless of risk of OHSS especially if the triggering of ovulation is made by GnRH agonist rather than by HCG.
◆Biosketch
Name: Khaled MAHMOUD
Date of birth: December, 10, 1954
Place of birth: Tunisia
Married and Father of 2 girls.
Scientific Degrees:
1985: Diploma in Obstetrics and Gynaecology University of Tunis
1985-1987: Diploma in the ART University of Marseille, France95

●Concurrent Symposium C-1.4

Training and ART practice at the Institute of Medicine of the Reproduction in Marseille France
Current position:
Founder and Head of the first Centre of IVF in Tunisia since April 1988
General Secretary of the Mediterranean Society of the Reproductive Medicine
Vice president of the Tunisian Society of Gynaecology and Obstetrics
Awards and distinction:
Medal of Merit as a pioneer of ART in Africa offered by the Federation of North African societies ofinfertility. Casablanca, Morocco decembre1997
Award of the best oral communication in the 4th MSRM congress
Member of the National Commission - within the Ministry for the Public health for the development and thestudy of the Assisted Reproductive Techniques (ART) in Tunisia.
Active participant in the preparation and writing of the law governing the ART at the Health Ministry in
Tunisia
Former Congress President of the 7th annual Meeting of Mediterranean Society for Reproductive Medicine
May 2008 Hammamet, Tunisia
Area of Special Interest and Accomplishments
My special areas of interest in Gynaecology and Obstetrics include infertility, both classical and endoscopic
surgery and high-risk pregnancy.
In the field of infertility I had the first babies born in Tunisia after GIFT (1989), ZIFT (1989), Embryo
reduction of Multiple IVF pregnancy (1992) and after ICSI (1996).
My works published or presented during the different meetings focused on ovarian stimulation, quality ofgametes and embryos, hysteroscopy and laparoscopy, male infertility, embryo implantation, genomics andmetabolic

96

●Concurrent Symposium C-2.1

Polycystic ovary syndrome is associated with metabolic abnormalities
in the purine mononucleotide pathway and tricarboxylic acid cycle
Makio Shozu1, Guiwne Wang1, Tomoya Segawa2,
Shokichi Teramoto2, Hiroshi Ishikawa1
1
Department of Reproductive Medicine, Graduate School of Medicine,
Chiba University, 2Shimbashi Yume Clinic
Introduction: Obesity associated with insulin resistance is a characteristic metabolic abnormality ofpolycystic ovary syndrome (PCOS), but is not the only feature of the metabolic status of such patients. To
investigate alternative therapeutic targets for PCOS, we analyzed the metabolomic profiles of PCOS patients.
Materials and methods: Plasma was obtained from women with PCOS (n = 5) presenting with obesity andinsulin resistance and women (n = 5) with regular menstruation presenting with simple obesity (BMI < 27).
Metabolites in plasma samples were analyzed using capillary electrophoresis followed by time-of-flight mass
spectrometry. Each metabolite was identified based on the measured variables and migration time using the
Kyoto Encyclopedia of Genes and Genomes database. Differences were considered significant when p < 0.05.
Results: The most striking difference was found in the levels of purine mononucleotides. There wereincreased levels of ATP, ADP, AMP, GDP, and IMP (the key intermediates of purine synthesis) in the PCOS
samples. In cells, levels of purine nucleotides are determined by equilibrium along the de novo synthesis
pathway, recycling pathway (salvage pathway), and catabolic pathway. In PCOS, the level of hypoxanthine, a
key intermediate of both the salvage and catabolic pathways, was not different from that of the control
subjects, suggesting that enhancement of de novo synthesis is responsible for the increase. In contrast to thepurine nucleotide levels, the level of pyrimidine mononucleotides was unchanged. The next striking difference
was the levels of intermediates of the tricarboxylic acid (TCA) cycle. Succinic acid, pyruvic acid, oxoglutaricacid, malic acid, and isocitric acid, but not citric acid, were increased in PCOS compared to the control
subjects.
Conclusion: PCOS is obviously associated with systemic changes in purine and TCA cycle metabolites,
which would not be a consequence of co-existing obesity. These metabolic abnormalities might be a new
therapeutic target of PCOS.
◆Biosketch
1981:
Graduated Kanazawa University, School of Medicine
1982-1986: Graduated Ph.D. program of Postgraduate School, School of Medicine, Kanazawa University
1986-1989: Department of Obstetrics and Gynecology, Fukui Prefecture Hospital
1990-1999: Lecture, allied Health Science School, Kanazawa University
1995-1998: Senior fellow, Cecil and Ida Green Center for Reproductive Medicine, University of Texas at
Dallas (Professor Evan Simpson).
1999-2005: Associated Professor, Center for Fetal and Maternal Medicine, Kanazawa University
Dec. 2005-present: Professor and Chairman, Reproductive Medicine, Chiba University.

97

●Concurrent Symposium C-2.2

How to get pregnancy in women with PCOS
Chii-Ruey Tzeng
Taipei Medical University
Polycystic ovary syndrome (PCOS) is an extremely common disorder in women of
reproductive age. Diagnosis of PCOS is principally based on clinical and physicalfindings. Diagnostic criteria and PCOS definitions used by clinicians and researchers are almost as
heterogeneous as the syndrome. Of those diagnosed with PCOS using the 2003 Rotterdam criteria, 61%
fulfilled 1990 NIH criteria for unexplained hyperandrogenic chronic anovulation. The patient populations with
the new phenotypes had less severe ovulatory dysfunction and less androgen excess than patients diagnosed
using the 1990 NIH criteria. These findings might be common across all female populations with PCOS,
whether in Oriental or Occidental countries. Data for clinical hyperandrogenism indicated that the prevalenceof hirsutism in Taiwanese PCOS women is lower than that for Caucasians/Western women.The extent of
metabolic abnormalities in women with PCOS may vary with phenotype, age and ethnicity. Obesity
represents a major risk factor for metabolic syndrome and insulin resistance. Approximately 40-50% of all
women with PCOS are overweight or obese. Obese subjects with PCOS had a higher risk of developing
oligomenorrhea, amenorrhea and biochemical hyperandrogenemia than non-obese women with PCOS.
Moreover, obese women with PCOS had significantly more severe insulin resistance, lower serum LH levels,and lower LH-to-FSH ratios than non-obese women with PCOS. PCOS women in Taiwan presented withhigher LH-to-FSH ratio and lower insulin resistance than PCOS women in Western Countries. However, theaverage body mass index (BMI) was significantly lower in Taiwanese PCOS women than Western women,
which might partially explain the difference between these two populations in terms of clinical andbiochemical presentations.The treatment of infertile women with PCOS is surrounded by many controversies.
Recently consensus meeting of ESHRE/ASRM are recommended as following:1.First-line treatment forovulation induction remains the anti-estrogen clomiphene citrate (CC).2.Second-line intervention is either
exogenous gonadotropins or laparoscopic ovarian surgery (LOS).3.Third-line treatment is in vitro fertilization(IVF).(The Thessaloniki ESHRE/ASRM-Sponsored PCOS Consensus Workshop Group. Consensus on
infertility treatment related to polycystic ovary syndrome, Fertil Steril 2008)Retrospectively reviewed ourclinical data, women with PCOS had excellent pregnancy outcome in Reproductive Medicine center of Taipei
Medical University Hospital.
◆Biosketch
Professor Tzeng is currently Dean of the College of Medicine at Taipei Medical University since 2004, andChairman of Ob/Gyn at TMU since 1992. He graduated from the School of Medicine at TMU in 1976 andobtained a MPH degree from Harvard School of Public Health from 1980-1981. He completed his fellowshiptraining in Reproductive Endocrinology and Fertility from 1981-1983 in the Department of Ob/Gyn at
Brigham & Women's Hospital, Harvard Medical School. He has undertaken two projects; the first test-tube
baby in 1985 and the first mitochondria transfer in 2002. He and his group have received the Prize Poster
Award at the annual meeting of ESHRE in 2001, 2003 and 2007 respectively. He received 2009 Gold Medalfor the invention of diagnosis method of endometriosis by detecting biochemical markers and the STARAward from ASRM in 2010. He served as President of the TSRM from 1996-1998, and PRSFS since 2008.
He was on the board member of the International Ovarian Conference and ISFP.

98

●Concurrent Symposium C-2.3

New criteria of diagnosis and treatment in polycystic ovary syndrome(PCOS) in Japan
Toshiro Kubota
Comprehensive Reproductive Medicine, Graduate School,
Tokyo Medical and Dental University
Polycystic ovary syndrome (PCOS) which is the most frequent endocrine disorder in women of reproductive
age could present a variety of clinical pictures. In Japan, compared to other countries, the usual clinicalpresentation of PCOS is slightly different - with less frequently encountered cases of hyperandrogenism therefore established European or US guidelines are clinically less useful. In 2006 the Japanese Society of
Obstetrics and Gynecology (JSOG) has proposed new, revised diagnostic criteria that in the future could also
be valued internationally. Diagnostic criteria are based on three main features: 1) cycle irregularities; 2)
polycystic changes in the ovary by ultrasonography; 3) endocrine anomalies (LH or androgen hypersecretion).
Based on the above-mentioned diagnostic criteria in 2008, the JSOG has also proposed revised treatment
guidelines summarized as follows:
1. PCOS patients wishing to have children: if obese (BM>25 kg/m2), as a first option, weight loss andexercise is recommended. Non-obese patients or those obese women who do not ovulate after lifestylechanges are submitted to ovulation induction therapy with clomiphene citrate. Obese clomiphene-resistantpatients who have impaired glucose tolerance or insulin resistance are treated with a combination of
metformin and clomiphene citrate. If the previously mentioned treatments options are unsuccessful, ovulationinduction with exogenous gonadotropin therapy or laparoscopic ovarian drilling (LOD) is recommended.
During ovulation induction a chronic, low-dose, step-down regimen is recommended with careful monitoringin order to reduce the risk of ovarian hyperstimulation syndrome (OHSS) and multiple pregnancies.
Alternatively with LOD high success pregnancy rates of around 60% are expected with a low risk of multiple
pregnancies. If ovulation induction is unsuccessful, in vitro fertilization (IVF) treatment is indicated. During
ovarian stimulation for IVF, FSH-only preparations are used. In high OHSS-risk patients, systematic embryo
freezing and subsequent frozen embryo transfer cycles are recommended.
2. PCOS patients not desiring for children: in non-obese, anovulatory patients, pharmacological treatments
such as Holmström, Kaufmann regimens or low-dose oral anticonceptives are used to induce regularwithdrawal bleedings which are especially important for preventing endometrial hyperplasia and endometrial
cancer.
These new diagnostic and treatment guidelines hopefully will contribute to an improved care of PCOS
patients in Japan.
◆Biosketch
1975
Graduated from Tokyo Medical and Dental University, Faculty of Medicine
1982-1991
Tokyo Medical & Dental University, Research Assistant
1988-1989
University of Virginia(U.S.A.), Research Fellow
1991-1996
Tokyo Medical & Dental University, Lecturer
1996-2006
Tokyo Medical & Dental University, Associate Professor
2006Tokyo Medical & Dental University, Professor

99

●Concurrent Symposium C-2.5

PCO and transvaginal hydrolaparoscopic ovarian drilling
Hiroaki Shibahara, M.D., Ph.D.
Department of Obstetrics and Gynecology, School of Medicine,
Jichi Medical University, Tochigi, Japan
Polycystic ovary syndrome (PCOS) is a syndrome of ovarian dysfunction showing
cardinal features of hyperandrogenism and polycystic ovarian morphology. It is one of the most common
reproductive endocrine disorders in young adult women, showing clinical signs of menstrual disorder,
anovulation, hirsutism, acne, and obesity. Frequently, this group of patients present with infertility due tochronic oligoovulation or anovulation. In cases that do not respond to medical induction of ovulation, ovarian
surgery such as laparoscopic ovarian drilling (LOD) by the trans-abdominal approach has been widely used toinduce ovulation in PCOS women after failure of treatment with clomiphene citrate. So far, many authors
have reported high rates of ovulation (~ 80%) and pregnancy (~ 60%) following LOD.
In 1998, transvaginal hydrolaparoscopy (THL) was introduced as the first line procedure in the exploration of
the adnexal structures in infertile women1). Because of the advantages of THL, including accurate inspectionof adnexal structures without manipulation, it became clear that THL is a less traumatic and more suitable
outpatient procedure than diagnostic laparoscopy. The risks of a general anesthetic are avoided, and there isless chance of trauma to major vessels2). Therefore, we have been performing THL for a diagnostic
laparoscopy on infertile women based on any of the following four indications3-6): i) tubal obstruction and/or
peritubal adhesion is suggested by hysterosalpingography, ii) serum antibody against C. trachomatis ispositive, iii) diagnosis of early-stage endometriosis, and iv) unexplained infertility. Our group also reported
the application and usefulness of THL for transvaginal salpingoscopy7).
Although THL was initially developed as a method for diagnostic laparoscopy, THL has more recently been
performed for operative laparoscopy, especially for ovarian drilling in women with PCOS8). This novel
technique, transvaginal hydrolaparoscopic ovarian drilling (THLOD) using a laser or bipolar electrosurgery,appears to be an effective minimally invasive procedure to induce ovulation in women with PCOS9-13). In areport, the cumulative pregnancy rate was 60 % for spontaneous and stimulated cycles, with 40% imputed todrilling alone. It was also reported that there were no complications.
The usefulness of diagnostic and therapeutic THL for the purpose of infertility investigation and treatment
will be presented.
◆Biosketch
Hiroaki Shibahara, M.D., Ph.D.
Professor, Department of Obstetrics and Gynecology, School of Medicine, Jichi Medical University, andCenter for Reproductive Medicine, Jichi Medical University Hospital, 3311-1 Yakushiji, Shimotsuke, Tochigi
329-0498, Japan
Education :Graduate:
Medical Doctor:
Ph.D.:

1984 Kochi Medical School (Nangoku, Kochi, Japan)
1984
1994 Department of Obstetrics and Gynecology, Hyogo College of Medicine(Nishinomiya, Hyogo, Japan)

100

●Concurrent Symposium C-2.5

Awards:1995: Hyogo Prefectural Society of Obstetrics and Gynecology
1998: Japan Society of Fertilization and Implantation
2003: Japan Society of Fertility and Sterility
Roles in the Medical Societies:
International
1) Councilor,
American Society of Reproductive Immunology
Domestic
1) Chair of the annual meeting
2007: the 5th annual meeting of the Japan Association of Psychological Counseling for
Reproductive Medicine
2011: the 6th annual meeting of the Japan Society for Reproductive Regeneration
2) Exective Board Members
Japan Society of Fertilization and Implantation
Japan Society of Mammalian Ova Research
Japan Association of Psychological Counseling for Reproductive Medicine
Japan Society for Reproductive Regeneration
3) Board Members
Japan Society of Gynecologic and Obstetric Endoscopy
Japan Society for Reproductive Immunology
Japan Society of Andrology
4) Ethics Committee
Japan Society for Reproductive Medicine
5) Editorial Board:
Reproductive Medicine and Biology (Editor-in-associate chief, Reviewing editor)
Current Women‘s Health Reviews
Open Women Health Journal

101

●Concurrent Symposium C-3.1

Available tools to personalize ovarian stimulation-Dose
Prediction/Protocol selection
Bruno Lunenfeld
Faculty of Life Sciences, Bar-Ilan University, ISRAEL
The response to ovarian stimulation is a key element and a major determinant in theoutcome of ART procedures. The response will depend on two variables: the dose of stimulant administeredand the protocol used and, the patient‘s response, depending on previous response, biological age (ovarian
reserve), as well as genetic variants, like the FSH and LH receptor and mutations of FSH and LH bythemselves. Age, antral follicle count and anti-Müllerian hormone seem to be the best indicators to predict
initial dose and protocol. Inhibin on day 3 of treatment will permit protocol assessment and dose correction.
Oestrogen and follicle count and size on day 6 will permit protocol adjustment adding LH, increasing or
decreasing the amount of FSH. Follicle count and oestradiol levels on the last day of treatment will help toavoid hyperstimulation by permitting the choice of agent to be used for ovulation induction, hCG, LH or
GnRH agonist, or freezing of the embryos◆Biosketch
Bruno Lunenfeld, MD, PhD, FRCOG, FACOG (hons), POGS (hons)
Born in Vienna 1927, married +2
Bruno Lunenfeld is Professor Emeritus at the Faculty of Life Sciences, Bar-Ilan University, and Ramat Gan,
Israel. He is also President of the International Society for the Study of the Aging Male, General Secretary of
The Asian-Pacific Initiative on Reproductive Endocrinology, Treasurer of the International Society of
Gynaecological Endocrinology and Editor of The Aging Male.
He graduated from the medical School in Geneva Switzerland. Following his post graduate work in Geneva hejoined the Weitzman Institute as senior Scientist, and then headed the WHO International Reference Center
for Fertility Promoting Drugs and acted as a Consultant and member of Expert Committees at the World
Health Organization.
Professor Lunenfeld is best known for his pioneering work in human reproduction. After describing theclinical use of hMG in men and women in 1960, his group was the first to achieve a pregnancy with hMG in
1961, demonstrated binding of hCG to LH receptors in theca cells in 1967 and were the first to induce
ovulation followed by pregnancy with GnRH in an hMG-stimulated cycle in 1975. His research interests
pertain to the physiology and pathology of male and female reproduction and include stimulators, modulatorsand regulators of sex steroids, spermatogenesis, spermiogenesis, folliculogenesis and ovulation, and themechanism of action of gonadotrophins, GnRH and growth factors on gene expression and steroidogenesis.
He was instrumental in the study of the aging male, was the founder of the International Society for the studyof the aging male (ISSAM. He organized in collaboration with the World Health Organization the first,
second and third world Congress of the aging male in Geneva.
He published more than 044 papers 19 books including a textbook on Male and Female Infertility and one the―first textbooks on Men‘s Health. He gave more than 700 invited lectures and chaired or co-chaired more102

●Concurrent Symposium C-3.1

than 100 sessions at national/international Meetings. He trained more than 200 local and foreign physicians
and supervised more than 50 MD, MSC, and Ph.D. Students
Professor Lunenfeld has received several distinguished international awards for his scientific achievements,
including the special recognition award of the United States Public Health Service for his outstanding
contribution in the promotion of health from the Surgeon General (1983), the Verdienstkreuz 1 Klasse – oneof Germany‘s highest honours – signed by the President of Germany, Roman Herzog (1995), the BertarelliFoundation Award for his lifetime achievements in women‘s health (2002) and the World Fertility AwarenessMonth Lifetime Achievement Award (2005).
Professor Lunenfeld is honorary member of European Society of Human Reproduction and Embryology(ESHRE), the International Federation of Fertility Societies (IFFS, German, Polish and Italian Society of
Gynecology and Obstetrics, German Endocrine Society of Reproduction, Austrian Society of Fertility and
Sterility, and the Asian Society of Andrology

103

●Concurrent Symposium C-3.2

Advances in ultrasound monitoring of IVF cycles by automated volume
measurement
Baris Ata1,3, Ayse Seyhan1, Hai-Ying Chen2, Alper Mumcu2,
Seang Lin Tan1,2
1
Dept. Of Obstetrics and Gynecology, McGill University, Montreal,
Canada2
Montreal Reproductive Centre, Montreal Canada
3
Dept. of Obstetrics and Gynecology, Uludag University, Bursa, Turkey
Accurate assessment of follicular size is crucial for good IVF treatment outcome. A major problem hitherto
has been large inter- and intra-observer variation in follicular measurements with conventional 2 dimensional(2D) ultrasound (US). Sonography-based Automated Volume Count (SonoAVC) is a new three-dimensional(3D) US technology, which automatically identifies and quantitatively analyses hypoechoic structures within avolume of interest. It generates a set of measurements including the mean diameter (MFD) and a
volume-based diameter (d (V)) for each hypoechoic structure in the scanned volume. Although SonoAVC wasdesigned for follicle measurement in the IVF setting, it has also been used to conduct volumetricmeasurements of embryonic or fetal structures.
Several studies have evaluated the accuracy of SonoAVC measurements and compared with actual follicularvolume or other validated methods of US volume measurements under a variety of conditions. In the IVF
setting, SonoAVC follicular measurement results are in very good agreement with conventional real time 2D
follicle measurements, actual follicular volume, and follicular measurements by VOCAL. SonoAVC results
maintain accuracy across different follicular sizes above 10mm. We have reported that the mean difference
between the size of the leading follicle on hCG day as measured by SonoAVC or conventional 2D US is lessthan 1mm. Using conventional hCG criteria for oocyte collection, exclusive use of SonoAVC for follicle
tracking does not improve or worsen clinical outcome as shown in a randomized controlled trial. The
technique seems to provide reliable and highly reproducible results. Automated measurements take less timethan manual measurements. Time savings can increase up to 5 minutes per patient. Most importantly,
SonoAVC measurements are highly reproducible. We have reported intracluster correlation coefficientsexceeding 0.8 for 4 different operators analyzing same volumes with SonoAVC. These results suggest very
low inter-observer variability. This is especially important for busy IVF programs where a patient can bescanned by different sonographers during the course of one IVF cycle. SonoAVC can facilitate remotemonitoring of women undergoing ovarian stimulation.
During early pregnancy scans, such as viability scans, SonoAVC allows accurate volume measurements ofirregular structures such as the embryo. It has been used to measure embryonic volume which can be a bettermeasure of embryonic growth than traditional crown rump length measurements. It has also been used toassess fetal heart and cardiac blood flow, volumes of cerebral ventricles. Again, SonoAVC yields accurate
results in shorter time compared other validated methods such as VOCAL, and makes it possible to introduce
these measurements into clinical practice. This new technology is expected to play a more important role in
the future of our practice.
◆Biosketch
Dr.Baris ATA,
Age 37
M.D. Istanbul University 1998
Residency Ob&Gyn Istanbul 2003

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●Concurrent Symposium C-3.2

Assisted Reproductive Technologies certification with Prof. Bulent Urman at the American Hospital of
Istanbul 2006
Attending Physician at the American Hospital of Istanbul IVF Unit 2006 – 2008
M.Sc. in Clinical Trials from the London School of Hygiene and Tropical Medicine, 2007 - 2010
REI Clinical Fellowship at McGill University, Montreal 2008 – 2010
REI Research Fellowship at McGill University, Montreal 2010 – 2011
Director of the Assisted Reproduction Unit of Uludag University, Bursa, Turkey Jan 2011 – ongoing
References: Prof. Seang Lin Tan and Prof. Victor Gomel
Authored – co-authored 35 papers indexed by SCI and 18 book chapters.

105

●Concurrent Symposium C-3.3

Volumetric measurements of the follicular development
Maximilian Murtinger
IVF Center Prof. Zech – Bregenz
Follicle tracking is commonly employed to assess the response to ovarian stimulation.
In the majority of cases two dimensional transvaginal ultrasound (2D) is performed tomonitor follicle growth and to determine the optimal time for administering human chorionic gonatotrophin(hCG) to trigger oocyte maturation. The presence of numerous follicles of different sizes and shapes makes
such an assessment much harder as the observer has to identify each and every follicle individually andmeasure it only once. The reliability and validity of such measurements are likely to reduce as the number offollicles increase. 2D only provides an approximation of the real volume of follicles and therefore cannot be
used to guarantee standards for follicle tracking.
This presentation will give an overview for the use of three dimensional ultrasound (3D) techniques combinedwith automated measurement of follicular size which might provide an objective, fast, valid and reliable
standard for such measurements in reproductive medicine.
◆Biosketch
Dr. Maximilian Murtinger received his MD at the University of Vienna in 1986. After specialization as a
trauma surgeon in 2003, he changed to the field of reproductive medicine and endocrinology and finished his
specialty in OB/GYN. since 2010.His current interest is in ultrasound techniques in reproductive medicine,especially related to follicular measurements in stimulated cycles and in establishing clinical standards related
to the quality criteria set by the European Union directive on tissues and cells (2004/23/EC). In addition, Dr.
Murtinger is developing requirements for software used in obstetrics and gynecology, especially forreproductive medicine and endocrinology. He is Deputy Medical Director at the IVF Center Prof. Zech –Bregenz.

106

●Concurrent Symposium C-3.4

Ovarian stimulation and endometrial receptivity
Pak Chung Ho
Department of Obstetrics and Gynaecology, The University of Hong Kong
Introduction: Ovarian stimulation is now used routinely in most in-vitro fertilization(IVF) programs. There was controversy as to whether excessive stimulation and high oestradiol (E2) level
would be associated with a reduction in the endometrial receptivity and implantation rate of the embryos in
IVF treatment. In a retrospective analysis, we have shown that the embryo implantation rates and clinicalpregnancy rates were significantly reduced in IVF treatment cycles only when the serum E2 level was morethan 20,000 pmol/l. Previous differences in results may be due to the use of different levels of serum E2 todefine excessive responders. Therefore, we conducted a series of studies on the endometrium of women
undergoing ovarian stimulation.
Materials and methods: We conducted a series of studies on the morphometry, blood flow and gene
expression in the endometrium of women undergoing ovarian stimulation for assisted reproduction as well as
normal women in natural cycles. We also studied the effects of oestrogens on the attachment of trophoblastic
spheroids (from choriocarcinoma cell lines) to the endometrial cancer cell lines.
Results: In a morphometric study of the glands and stroma in endometrial biopsies of women in whom
embryo transfer could not be performed for a variety of reasons, we found that the glands and stroma inmoderate responders were in phase as in women in unstimulated ovulatory cycles. In excessive responders,
there was evidence of gland-stroma dys-synchrony. Doppler studies of the endometrial blood flow also
showed that there was a significantly higher percentage of excessive responders with no endometrial color
signals in the endometrium. Our results on the endometrial gene expression patterns in natural cycles,
moderate responders and excessive responders showed that there was a significant difference (more than 2fold) in the expression patterns of 411 genes among the 3 groups. A subsequent study showed that the gene
expression pattern of the excessive responders on day LH+7 was more similar to that of day LH+10 in natural
cycles, indicating that there was advancement in the endometrial development in excessive responders. Using
an in-vitro spheroid-endometrium attachment model, we also demonstrated that the attachment of spheroids toendometrial cancer cell lines was adversely affected by high levels of oestrogen.
Conclusion: In conclusion our series of studies showed that excessive response to ovarian stimulation willaffect the morphological development, the blood flow, gene expression patterns and receptivity of theendometrium. There is a need to avoid excessive ovarian stimulation in IVF programs.
◆Biosketch
Professor P.C. Ho is the Chair Professor of Obstetrics and Gynaecology, Director of Centre of Reproduction,
Development and Growth, and Associate Dean in Clinical Affairs in Faculty of Medicine, University of HongKong. He is in charge of the assisted reproduction program at the University of Hong Kong/Queen Mary
Hospital Centre of Assisted Reproduction and Embryology. His research interests include medical abortion,
emergency contraception, infertility and assisted reproduction. He has published more than 350 papers inpeer-reviewed journals. He is serving on many national and international professional bodies. He is currently
the President of Asia-Oceania Federation of Obstetrics and Gynaecology, and President of Family Planning
Association of Hong Kong.

107

●Concurrent Symposium C-3.5

Minimal ovarian stimulation (mini-IVF) for IVF utilizing vitrification andcryopreserved embryo transfer
John Zhang1, Lyndon Chang1, Yoshie Sone-Endo1,
Sherman Silber1,21
New Hope Fertility Center, New York, U.S.A.,
2
Infertility Center of St. Louis at Luke's Hospital, Missouri, U.S.A.
Gentle ovarian stimulation protocols, such as mini-IVF, have several potential advantages over conventional
IVF protocols, including less medication and fewer injections, producing fewer eggs, but eggs of higher
quality. The particular mild stimulation protocol called mini-IVF is described. This protocol requires a reliable
and cheap method for embryo cryopreservation such as vitrification, because of the negative impact ofclomiphene citrate on the endometrium and since cryopreserved embryo transfers with this protocol haveyielded much higher pregnancy rates than fresh transfers. In this series, patients were not denied treatment
based on their day-3 FSH value or ovarian reserve. Yet very acceptable pregnancy rates were achieved (20%for fresh embryo transfers and 41% for cryopreserved embryo transfers). These results strengthen theargument for a mini-IVF protocol and vitrification as an alternative to standard conventional IVF stimulation
protocols. Now a randomized control trial with cryopreserved single-embryo transfer is required.
◆Biosketch
Dr John Zhang graduated in 1984 from Zhejiang University School of Medicine in China. He was awarded his
PhD from Cambridge University for his IVF studies in 1991. In addition, he also has a Masters degree in maleinfertility from Birmingham University, UK. He is a reproductive endocrinology and infertility specialist
whose academic training includes residency training in obstetrics and gynecology and a fellowship in
Reproductive Endocrinology and Infertility in 2004 from New York University School of Medicine. Dr Zhang
is the director and founder of New Hope Fertility Center, an independent IVF facility in New York.

108

●Concurrent Symposium C-3.6

Poor responder patients undergoing IVF treatment
Svend Lindenberg Professor Dr.Med.
Copenhagen Fertility Center, Copenhagen
The definition of poor responder is still being debated although ESHRE 2011 has
added much better clarification to this issue.
The presentation will use evidence based methodology to elucidate several aspects of the possible medical
target points for poor responder patients. However very little new information has been generated during thelast 10 years and very little has added to the treatment modalities although several new drugs have been
introduced on the marked.
Specifically we present data on Growth Hormone and luteal phase modulation providing hope for better andmore efficient treatment for these patients.
However the general conclusion is that we still need a better understanding of the pathophysiology in poor
responder patients to improve treatments.
◆Biosketch
Professor dr. med Svend Lindenberg is a pioneer in IVF and has partcipated in the group of scientist
producing the first IVF baby in Denmark in 1982. Since then published more than 150 papers and given
numerous presentations in internatinal and national societies and holds several international patents in ART.
Today director for Copenhagen Fertility Center and Research Institution with focus on low stimualtion and
IVM.
Copenhagen Fertility Center: Copenhagen, Lygten 2c, DK-240 NV
In 2012 Congres President for the 2012 internationa Meeting in ISMAAR: www.ismaar2012.dk

109

●Concurrent Symposium C-3.7

Prediction and prevention of the ovarian hyperstimulation syndrome(OHSS) - an evidence-based approach
Hassan N. Sallam, MD, FRCOG, PhD (London)
Professor of Obstetrics and Gynaecology, the University of Alexandria,
Clinical and Scientific Director of the Alexandria Fertility Center, andDirector of the Alexandria Regional Center for Women‘s Health andDevelopment, Alexandria, Egypt
Ovarian hyperstimulation syndrome (OHSS) is an uncommon but serious iatrogenic complication of ovarian
stimulation occurring during the luteal phase or during early pregnancy. It is potentially fatal and is difficult topredict.
Prospective controlled studies have shown that good predictors include young age, the presence of polycystic
ovarian syndrome or polycystic ovaries on ultrasound, the measurement of the antral follicular count (AFD),
estradiol levels, insulin resistance, ovarian volume or antimullerian hormone (AMH) before starting ovulationinduction. Bad predictors include the measurement of body mass index (BMI), genetic predisposition, serum
vascular endothelial growth factor (VEGF), von Willebrand factor and perifollicular blood flow.
Similarly, randomized controlled trials (RCTs) have shown that primary prevention (before starting
HMG/FSH) can be achieved by giving FSH rather than HMG (without GnRHa), using the step-up rather thanthe conventional protocol, giving GnRH antagonists rather than agonists (but with a lower LBR), performing
IVM rather than IVF and using the sequential rather than the step down protocol. They have also shown thatthe alternate day protocol is comparable to the conventional protocol and that the sequential protocol iscomparable to the step-up protocol, while the superiority of either the step-up compared to step down protocol
needs further evaluation.
RCTs have also shown that secondary prevention can be achieved by triggering ovulation with GnRH agonists,
metformin administration, intravenous albumin, hydroxyethyl starch, cabergoline (for early OHSS) andlaparoscopic ovarian electrocautery. They have also shown that the following approaches are equivocal inpreventing OHSS: coasting versus unilateral oocyte aspiration and GnRH antagonists versus coasting, while
the following approaches await further evaluation: cancellation of the cycle, coasting, diminishing the dose of
HCG, embryo freezing and triggering with GnRHa + embryo freezing.
◆Biosketch
Hassan Nooman Sallam, MB, ChB, DGO, DrChO&G (Alex), FRCOG (England), PhD (London)
Hassan Nooman Sallam is professor and former head of the department of obstetrics and gynaecology inAlexandria University and clinical director of the Alexandria Fertility and Assisted Reproduction Center. He
is also the director of the Alexandria Regional Center for Women's Health and Development, a training andresearch center active in all areas of women‘s health based in Alexandria, Egypt. He is the former vice-dean
and director of research in Alexandria University Faculty of Medicine. In 1998, he served as the overseas
examiner to the Royal College of Obstetricians and Gynaecologists in London.
Besides the fellowship of the Royal College of Obstetricians and Gynaecologists (FRCOG), he holds a
doctorate degree in Obstetrics and Gynaecology from Alexandria University and a PhD in reproductive
biochemistry from the University of London. He has published more than 100 papers in national and110

●Concurrent Symposium C-3.7

international refereed journals in English and French as well as 3 books. He is the co-author of ―Infertility andAssisted Reproduction‖, the reference textbook published by Cambridge University Press.
Professor Sallam serves/served on the editorial boards of many journals including "Human Reproduction","Reproductive Biomedicine Online", "Middle East Fertility Society Journal" and ―Facts, Views and Visions in
OB/GYN‖ (Official Journal of the Flemish Society of Obstetrics and Gynecology) and is an ad hoc reviewer
for 11 other journals. He is a founding board member of the Middle East Fertility Society, based in Beirut,
Lebanon and the founding chairman and current president of the Mediterranean Society for Reproductive
Medicine, based in Florence, Italy.

111

●Concurrent Symposium C-3.8

What should we prefer for IVF-The Agonist or Antagonist?
Prof Zion Ben-Rafael
Tel Aviv, Israel
The answer seems clear. While the combination of GnRH-Antagonist for IVF iscontroversial, the use of GnRH-Agonist is widely accepted. Therefore, after two decades of experience and
200 publications one can't escape from the conclusion that for most clinician the Antagonist is a second choicereserved for special cases. Yet this clear cut preference by clinicians is not fully supported by the evidence.
Combining GnRH –analogs (agonists or antagonist) became the standard of care in all IVF treatments, with
the Agonist accounting for more than 2/3 of the treatments. GnRH antagonist with it direct –competitive
blocking action on the receptors to GnRH in the pituitary was suppose to be better than the old time agonist,
which is plugged by several side effects.
The advantages to the use of the antagonist which includes; Short treatment duration, Lower gonadotropins
requirement, Lower OHSS rate, Lack of severe hypoestrogenism and its consequences, Fitness to be used in
spontaneous cycles. The disadvantages include: A novel approach which is still under evaluation and needs
more optimization. If not given early enough, might fail to prevent high endogenous LH and most importantly
pregnancy rate per cycle is consistently lower than with the agonist. Most protocols allow minimal
programming; luteal phase support is also needed.
Conclusions: The GnRH agonist long luteal phase protocol should be the protocol of choice for all young IVF
patients. All other protocols of agonist or antagonist protocols should be reserved for special cases andsituations; apparently the best Antagonist protocol is the fixed daily dose protocol which should be preferredover the flexible or one dose protocol. Other Antagonist protocols should be developed to match the analog
results. Patients at higher risk for OHSS might enjoy a milder stimulation and lower overall risk with the
GnRH-Antagonist.
◆Biosketch
Prof. Zion Ben-Rafael,
Married + 3
Graduated from Tel Aviv University, Board certified in Israel and USA.
84-86, NIH Fellow in Reproductive Biology and Endocrinology, in Hospital of University of Pennsylvania,
Philadelphia.
Full Professor (1993), Associate Professor (1990), Tel Aviv University and Incumbent of the Tarnesby- Chairof Fertility regulation Tel Aviv University
Previously:
President, Israeli Menopause Association.
1997-2007 Chairmen, Department of OBGYN, Rabin Med Center.
1991-2007 Chairmen, Department of OBGYN, Golda-Hasharon.
1989-1991 Director, Katowitz Women's Health Center, Tel Aviv
Currently in charge of the IVF unit, Laniado hospital112

●Concurrent Symposium C-3.8

Founder and Chairman, COGI - World Congress on Controversies in Obstetrics, Gynecology and Infertility.
1999 - Prague, 2001 - Paris, 2002 - Washington, 2003 - Berlin, 2004 - Las Vegas, 2004 - Bangkok, 2005 Athens. 2007, Barcelona 2007 - Shangahi, 2008 - Paris, 2009 - Beijin, 2010 - Berlin, 2011 - Paris, 2011 Hainan china.
Founder and Chairman of the Mediterranean Congress 1993-96
Founder and Co-Chairman, Gender based medicine, World Congress Berlin 2006
Co-Founder World congress on controversies in Obesity, Diabetes and Hypertension 2006
Scientific Publications:
Total over 350 Scientific Publications; 280 original articles, 2 books, editor 12 congress proceedings

113

●Concurrent Symposium C-4.1

Evidence-based management of endometriosis-associated infertility
Hassan N. Sallam, MD, FRCOG, PhD (London)
Professor of Obstetrics and Gynaecology, the University of Alexandria,
Clinical and Scientific Director of the Alexandria Fertility Center, andDirector of the Alexandria Regional Center for Women‘s Health andDevelopment, Alexandria, Egypt
Various therapies have been used in the management of endometriosis-associated infertility. These include
surgical therapy, medical therapy, combined medical and surgical therapy, controlled ovarian
hyperstimulation as well as assisted reproductive techniques. Observational studies have shown that expectant
management for 18 months is associated with about 50% cumulative pregnancy rate in patients with stage I
and II endometriosis, while patients with severe degrees of endometriosis rarely become pregnant (Olive et al,1985). Surgical management of endometriosis includes ablation and/or resection of laparoscopic lesions aswell as the drainage of endometriomas. In a meta-analysis, Hughes et al found that surgical intervention in
general was associated with a higher pregnancy rate compared to either medical therapy or no therapy, but thestudies were heterogeneous making this conclusion doubtful (Hughes et al, 1993). In a study by Jones andSutton, drainage of endometriomas with ablation or resection of their walls was associated with a highpregnancy rate but there are no randomized trials to confirm these findings (Jones and Sutton, 2002). In
women with minimal-to-mild endometriosis, pregnancy rates following laparoscopic ablation and/or resection
were found to be significantly higher than in women who had diagnostic laparoscopy in a multi-center
randomized study conducted in North America (Marcoux et al, 1998), but these results could not be confirmed
in another multi-center study conducted in Italy (Parazzini, 1999). On the other hand, meta-analyses ofrandomised studies have shown that danazol, gestrinone, medroxyprogesterone acetate and GnRH agonists do
not enhance pregnancy rates over placebo or no treatment (Hughes et al, 1993). Similarly, the combination of
laparoscopic surgery and medical therapies did not improve pregnancy rates over danazol-only therapy (Yapet al, 2004). Controlled ovarian hyperstimulation combined with intrauterine insemination (COH+IUI)
improves the pregnancy rates significantly compared to expectant management (Tummon et al, 1997). Finally,
in-vitro fertilization (IVF) is associated with a high pregnancy rate in women with endometriosis, although
this is lower than in those with tubal factor infertility (Barnhart et al, 2002). In a recent meta-analysis, we haveshown that long-term (3 to 6 months) administration of GnRH agonists prior to IVF in these patients improves
the pregnancy rate significantly (Sallam et al, 2005).
◆Biosketch(For Professor Sallam‘s Biosketch, please refer to his abstract for Concurrent Symposium C-3.7.)

114

●Concurrent Symposium C-4.2

Surgery of Endometriosis in Infertility: State of the Art!
Issam Lebbi
Professor in gynaecology-obstetrics, Dream Center, 1002, Tunis, Tunisia
In the field of endometriosis associated infertility, we know since perhaps 15 years
now that:
1-The surgical treatment of the mild stages (stages 1 and 2) improve significantly the spontaneous pregnancies
rates (17.7 % to 31.7 %, ENDOCAN study, NEJM; 1997).
2-The surgical treatment of the moderate and severe stages (stages 3 and 4) can or should improve thespontaneous pregnancies rates (evidence level 3 by expert opinions).
The gold standard treatment of endometriosis-associated infertility is done by laparoscopic surgery. Therefore,
the place and the interest of this procedure in the management of the disease is highly discussed in the era of
ART and the main question is: how many laparoscopies should we perform in this group of patients to obtain
one pregnancy with regard to the physical, psychological, social and financial costs of laparoscopy?
Indeed, the place of laparoscopy in an integrated approach with ART techniques in the fields of endometriosis
associated infertility and unexplained infertility is not resolved yet.
Since five years now, surgical treatment for endometriosis associated infertility is ―gaining popularity
supported mostly by uncontrolled studies‖!
These studies reported a good pregnancies rate (55 to 65 %) with surgery after IVF failure, surgery as a firstline treatment and/or before IVF-ET and surgery before IVF-ET in severe stages and deep infiltrative
endometriosis.
In this lecture we will try to propose an integrated approach of the management of endometriosis associatedinfertility but as more controlled studies are needed in this field, we can‘t, in my opinion, talk about a state of
the art in the management of these situations.
◆Biosketch
Professor Issam Lebbi
Professor in gynaecology-obstetrics
Dream Center, 1002, Tunis; Tunisia
Polyclinique Alyssa, Les Berges du Lac, Tunis
Medical Director of IMBR (Institute of Medicine and Reproductive Biology)
Past-President of STGO (Tunisian Society of Gyn-Obs)
Past-President of FGOM (Mediterranean Federation of the National Societies of Gyn-Obs)
Member of the Board of Directors of ISIVF
Honorary Member if the CNOGF (French College of Gyn-Obs)

115

●Concurrent Symposium C-4.3

Surgical management of recurrent endometrioma: prior to IVF or not
Hiroshi Nabeshima, M.D., Ph.D.
Department of Obstetrics and Gynecology, Tohoku University Hospital
Since ovarian endometrioma is frequently diagnosed in women of reproductive age,laparoscopic excision of the endometrioma is performed for most cases. However,endometrioma frequently recurs even after repeated surgical procedures.
When, we perform the IVF for the patient with endometrioma, we wonder whether the endometrioma should
be removed or not before the egg collection, especially the endometrioma is recurrent one.
In this session, we show our data of recurrent endometrioma of IVF/ET patients and literature review at first.
After that, we show the laparoscopic minimal invasive technique for the removal of endometrioma by VIDEO,
and our surgical management to prevent postoperative ovarian dysfunction in IVF/ET patient.
◆Biosketch
Hiroshi Nabeshima, M.D., Ph.D.
EDUCATION
1997
M.D., Akita University School of Medicine
2004
Ph.D. (Dr. of Medical Science), Tohoku University Graduate School of Medicine.
2010
Attending to the Frontier In Reproduction course of Marine Biological Laboratory (Woods Hole,MA)
PROFESSIONAL TRAINING and EMPLOYMENT
2009- Present
Medical Staff and Researcher of Reproduction Medicine Unit in Obstetrics andGynecology, Tohoku University Hospital.
2005- 2009
Medical Staff in Obstetrics and Gynecology, Iwate Prefectural Iwai Hospital, Ichinoseki,
Iwate, Japan.
(2007-2008
Kurashiki Medical Center for laparoscopic fellow training)
2004- 2005
Medical Staff in Obstetrics and Gynecology, Iwate Prefectural Miyako Hospital, Miyako,
Iwate, Japan
2000- 2004
Resident and fellow in Department of Obstetrics and Gynecology, Tohoku University
Graduate School of Medicine, Sendai, Miyagi, Japan
1999-2000
Resident in Obstetrics and Gynecology, Sanuma Hospital, Toyoma, Miyagi, Japan
1997-1999
Resident in Obstetrics and Gynecology, Yuri Kumiai General Hospital, Yurihonjo, Akita,
Japan
1997
Passed the Examination of National Board
CERTIFICATION and LICENSURE
1997
Japanese Medical License Registration
2002
Diplomate, Japanese Society of Obstetrics and Gynecology, Certificate
2005
Technical Fellow of the Japan Society of Gynecological and Obstetric Endoscopy, Certification
2006
Technical Fellow of the Japan Society for Endoscopic Surgery, Certification
2009
Japan Society for Reproductive Medicine Board Certified Reproductive medicine specialist by Japan
Society for Reproductive Medicine, Certification

116

●Concurrent Symposium C-4.4

MANAGEMENT OF HYDROSALPINGES PRIOR TO IVF AND ET
Victor Gomel
Professor, Department of Obstetrics and Gynecology,
Faculty of Medicine, University of British Columbia,
Vancouver BC. Canada
We have been aware of the deleterious effects of hydrosalpinx on the outcome of IVF
treatment. This deleterious effect is more evident with large hydrosalpinges, visible at sonography. It has also
become evident that the major detrimental effect is due to a ―wash-out effect‖ owing to the passage of thecollected tubal fluid to the uterine cavity at the time, or soon after the transfer of the embryos to the uterine
cavity. This ―wash-out‖ may also occur sometime after transfer; as evidence suggests many embryos ascend
to the tube after transfer and eventually return to the uterus when the tube assumes a pro-uterine transport.
The following therapeutic options have been used to overcome this adverse effect:
1. Aspiration of the hydrosalpinx at the time of oocyte retrieval or embryo transfer.
2. Proximal occlusion of the tube.
3. Salpingectomy.
4. Salpingostomy.
Several studies reported on aspiration of the liquid contained in the hydrosalpinges. While some reportsignificantly higher pregnancy rates with aspiration others do not. This approach remains controversial.
Laparoscopic tubal occlusion has been shown to improve the odds of clinical pregnancy after meta-analysis.
However results for ongoing pregnancy, reported by only one trial, were not significant.
It has been demonstrated that salpingectomy before IVF, clearly improves the outcome of IVF treatment; thisis more evident with large hydrosalpinges, visible at sonography.
Salpingostomy as opposed to salpingectomy may be used in appropriately selected cases. The beneficial effectof this approach in IVF has been demonstrated in a small number of cases. Obviously salpingostomy, in
addition, offers the woman the potential of achieving a pregnancy naturally.
The place for each one of these treatment modalities and the selection of the appropriate option will be
discussed in detail.
◆Biosketch
Dr. Gomel holds the rank of Professor in the Department of Obstetrics and Gynecology, Faculty of Medicineof the University of British Columbia. He served as Chairperson of his Department for fifteen years, during
which the Department was greatly expanded and attained international recognition.
Dr. Gomel‘s special research interests lie in the fields of gynecologic surgery and reproductive medicine. In
the field of human reproduction, his work included infertility associated with tubal and peritoneal causes,
fertility control, reproductive surgery, operative laparoscopy and in vitro fertilization. His other researchinterests include reproductive physiology and formation and prevention of peritoneal adhesions.
He is internationally known for his pioneering work in both microsurgery and operative laparoscopy. He was
responsible for the introduction of laparoscopy in Canada; he was the first to demonstrate the efficacy oflaparoscopic fertility-promoting procedures and one of the first to use laparoscopic surgical access for othergynecologic procedures.
For nearly four decades, he has been involved in work to better understand the tubal and peritoneal factors thatcause infertility, and to improve the surgical techniques to correct them. His experimental work was designed

117

●Concurrent Symposium C-4.4

to elucidate the function of the fallopian tube. Microsurgical techniques were employed to alter the structureof the fallopian tube and thus gain understanding of the function of its various parts. Another aspect of his
animal work was designed to better understand the formation of peritoneal adhesions. Various techniques andnumerous substances were used in the attempt to diminish or prevent the formation of peritoneal adhesions.
This work was carried out with the collaboration of members and various trainees (fellows) in his Department.
These efforts led to the development and introduction of microsurgical techniques in gynecology, which
produce significantly better results. These techniques have gained international acceptance and recognition.
He established an in vitro fertilization (IVF) program in Vancouver in 1981. Under his leadership thisprogram was the first to achieve success, resulting in the birth (on December 25, 1983) of the first IVF baby in
Canada.
He was responsible for the establishment of a masters and PhD. Program ―Reproductive and Developmental
Sciences‖, within his Department, which has been operative since 1984; at present this program has more than
40 graduate students. He was also instrumental in the creation of the ―Women‘s Health Centre,‖ which was
the first such tertiary facility in Canada, and has been operational since early 1992.
In addition to medical students and residents he has trained, in Vancouver, post doctoral fellows from all over
the World, many of whom currently hold important positions, in Canada and in their own countries.
Dr. Gomel authored a long list of scientific articles published in prestigious international journals. He wrote
numerous book chapters, and authored and edited several books. His authored books, among others, include―Gynecology: A Practical Approach‖, co-authored with Malcolm Munro and Timothy Rowe (Williams andWilkins 1990), a problem oriented text designed for medical students, interns, junior residents and generalpractitioners. Published in 1983 (Little Brown & Co.), his book ―Microsurgery in Female Infertility‖ isconsidered a classic. His book ―Diagnostic and Operative Gynecologic Laparoscopy‖ (Mosby 1995) was
highly acclaimed. In addition he has been the editor or co-editor of several other books including ―Female
Genital Prolapse and Urinary Incontinence‖ (Informa 2008). His most recent book is ―Reconstructive andReproductive Surgery in Gynecology‖ (Informa 2010).
He is on the editorial boards of numerous important international scientific journals. He has been on the Boardof Trustees and Executive Boards of many national and international scientific societies, including among
others, the American Society of Reproductive Medicine (previously AFS), the American Association of
Gynecologic Endoscopists (AAGL) and the International Society of Gynecologic Endoscopy. He served aspresident several societies, including the Society of Reproductive Surgeons, the AAGL, and the CanadianFertility and Andrology Society. He is currently the Vice president of the Peritoneum and Surgery Society andthe Vice President of the International Society of In Vitro Fertilization (ISIVF).
He has received honorary memberships and awards of excellence from numerous international scientificsocieties and universities, including his own Faculty of Medicine, in recognition of his pioneering work in
reproductive medicine and gynecologic surgery. Professor Gomel was also the recipient, of the prestigious
distinction of Chevalier of the order of Légion d‘Honneur, awarded by Jacques Chirac, President of France in
2003. In 2008, he was elected ―Fellow‖ to the World Academy of Art and Science, and in 2009 he received
the degree of Doctor of Science, Honoris causa, from the Simon Fraser University.

118

●Concurrent Symposium C-5.1

Surgery of intramural leiomyomas in infertility patients
Jean-Bernard Dubuisson, Victor Gomel
Department of Obstetrics and Gynecology (HUG, Genève)
Uterine myomas are the type of pelvic tumour found most frequently, and thisraises the problem during everyday practice of how to manage them. This isparticularly clear in infertility patients. When uterine myomas are diagnosed this does not necessarily meansurgery is required. The development of endoscopic procedures (hysteroscopy and operative laparoscopy), ofembolization and IVF procedures have modified the over the past few years the operating indications foruterine leiomyomas. Only complicated or large intra mural leiomyomas distorting the uterine cavity needsurgical treatment together with those giving rise to symptoms which do not respond to properly conducted
medical treatment. Technical advances mean that surgical treatment today for uterine myomas can for certain
indications be carried out by operative laparoscopy.
The two most crucial elements for infertility patients in laparoscopic myomectomy are the quality of theuterine suture and the risk of adhesions.
The problem of the quality of the uterine suture is above all important for women desiring pregnancy. The risk
is low and depends mostly of the quality of the surgery.
The second problem with laparoscopic myomectomy is the risk of adhesions. Once again the risk is not
specific to laparoscopic surgery, for adhesions have been observed in 80% of the cases after myomectomy via
laparotomy. In our experience the rate of adhesions we observed after laparoscopic myomectomy is less.
All these elements enable us to stress that just as for any surgical technique, once it has been shown to befeasible the most important element is the indication for surgery. We believe that laparoscopic surgery is
indicated for intramural leiomyomas, only in selected indications that will be discussed.
◆Biosketch
Professeur Jean-Bernard DUBUISSON
Date of Birth: 10th May, 1946 in Paris (France), Gynaecological Surgeon
Head department of obstetrics and gynecology (HUG, Genève) since 2004
Head unit of gynecology (HUG, Genève) since 2003
Ordinary Professor of the University of Geneva since 2003
Docteur Honoris Causa, Faculté d‘Athènes, septembre 2005
Professor of the University of Paris (Université Paris V, Centre Hospitalier Universitaire Cochin Port-Royal,
Paris) 1980 - 2003
Head Department of Gynecology (Hôpital Baudelocque, Centre Hospitalier Cochin - Paris) 1993 – 2003
DIPLOMAS :Interne des Hôpitaux Paris (1970), Medical Doctor (thesis of Medicine Paris 1975), General Surgeon (1977), Obstetrician and Gynaecologist (1978)
SCIENTIFIC SOCIETIES :France :Member of the French Commission of the Universities 1998 - 2003
Director of University Certificate "Etudes Relatives à la Stérilité et aux Troubles de la Reproduction" (Paris)
1988 - 2003

119

●Concurrent Symposium C-5.1

Director of University Certificate "Applications de l'Endoscopie en Gynécologie" (Paris) 1992 - 2003
President of FIV-NAT Association 1987-1989
Vice-President of the French Society of Gynaecological Endoscopy since 1989 (SFEG)
Member of the French Academy of Surgery 1995-2006
Europe :Member of the Executive Board of the European Society of Gynaecological Endoscopy (1993-1999)
Member of the Editorial Board of HUMAN REPRODUCTION, Coordinator of the Special Interest Group of
Reproductive Surgery of ESHRE (European Society of Human Reproduction and Embryology)
USA :Member of the Board of Trustees of the American Association of Gynecologic Laparoscopists (1998-1999),
member of the Editorial Board of AAGL (1999-2005) and International Advisor of AAGL (since 2006)
Switzerland :Member of the Swiss Society of gynecology & obstetrics (since 2003)
Member of the Management Committee of Swissendos (since 2003)
PUBLICATIONS: 200 publications in international journals, 150 publications in French journals, 100
publications for teaching, 100 chapters in books, 4 books :. Coelioscopie et cancérologie en gynécologie
J.B. Dubuisson, C. Chapron, J. Bouquet de la Jolinière
Editions Arnette 1993
. Les fibromes utérins
J.B. Dubuisson, C. Chapron, J. Bouquet de la Jolinière
Editions Arnette 1994
. L'endométriose
J.B. Dubuisson, C. Chapron, J. Bouquet de la Jolinière
Editions Arnette 1995
(traduction anglaise : endometriosis). La douleur en gynécologie
C. Chapron, D. Benhamou, J. Belaisch Allart, J.B. Dubuisson
Editions Arnette 1996 sous presse
These publications concern:
Microsurgery in infertility (1978 – 1988)
IVF (natural cycles, CIVETE) (1985 – 1990)
Operative laparoscopy (1990 – 2006):- tubal infertility (laparoscopic salpingostomy, laparoscopic tubal anastomosis),- myomas (laparoscopic myomectomy, laparoscopic hysterectomy),- endometriosis (laparoscopic treatment of endometriosis of the bladder, deep endometriosis),- genital prolapse (new procedures of laparoscopic lateral colpo-uterine suspension with or without
meshes),- tubal pregnancy (laparoscopic salpingectomy, management)120

●Concurrent Symposium C-5.2

Myomectomy during pregnancy
Yuji Hiramatsu
Department of Obstetrics & Gynecology, Okayama University Graduate
School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama,Japan
There are two kinds of treatment for pregnant women complicated with uterine myomas. One is conservative
therapy while the other is myomectomy during pregnancy.
Today, I would like to talk about a myomectomy during pregnancy. This operation is still prohibited
according to most textbooks. About 20 years ago, a pregnant women experienced deep vein thrombosis andpulmonary embolism with a 15 cm myoma. She had a D/C and was treated in the ICU for 2 weeks. We also
experienced two mid-trimester fetal deaths in other patients. Thereafter, we began to perform myomectomies
during pregnancy.
There are some merits and demerits in both therapies. In conservative therapy, there is no risk of abortion
caused by operation, but many other complications may occur during pregnancy which may result in longhospitalization, birth troubles and the incidence of cesarean section may increase. On the other hand, with amyomectomy, there is a risk of a spontaneous abortion after the operation; however, these symptoms
disappear in 87% of all such patients.
We have so far performed 92 myomectomies during pregnancy. The size of myomas was 4-25cm and thenumber of enuclated myomas was 1 - 25. The indications of myomectomy include, 1) symptoms of threatened
abortion do not disappear in spite of conservative therapy, 2) a rapidly enlarging myoma, 3) a past history of aspontaneous abortion or IUFD which might have been caused by the myoma, 4) myomas which may cause
complications during pregnancy, 5) myomas which cause recurrent degeneration and intense pain.
The contra-indications are 1) the margin of the myoma is not clear (adenomyosis), 2) a posterior cervical
myoma.
The most common symptoms are abdominal pain, genital bleeding and rapidly enlarging myomas. Somepatients present with more than 2 symptoms.
We followed up 83 patients, excluding 9 that experienced spontaneous abortions. The symptoms disappearedin 87% of the patients and they could enjoy as same lifestyle as other normal pregnant women. Thirteen
percent of the patients required some therapy for a threatened spontaneous abortion or preterm labor.
Seventy-nine cases (87.3%) had term delivery and 34 cases (43%) of those had a vaginal delivery. Four cases(2.5%) were preterm delivery and 9 cases (10.1%) had a spontaneous abortion. However, one IUFD occurred
after 5 weeks and one placental abruption occurred 9 weeks after the operation. In results, 7 cases (7.6%)
experienced abortions caused by a myomectomy.
A myomectomy during pregnancy is considered to a safe and beneficial treatment when performed by doctors
who are Knowledgeable and experienced in using the proper techniques for a myomectomy.

121

●Concurrent Symposium C-5.2◆Biosketch
Education
1971-1977 Okayama University Medical School
1981-1985 Graduate School of Okayama University Medical School
Academic Appointments:
1989-1995 Instructor of Okayama University Hospital
1995-1999 Assistant Professor of Okayama University Hospital
1999-2003 Associate Professor of Okayama University
2000
Visiting Associate Professor of Harvard Medical School, Joslin Diabetes Center
2003Professor & Chairman of Okayama University Graduate School, Department of Obstetrics &Gynecology
Major Committee Assignment:
Japan Society of Obstetrics and Gynecology
The Japan Society of Diabetes and pregnancy
The Japan Society of Gynecologic Oncology
Japan Society of Gynecological and Obstetrical Surgery
Memberships:
American Diabetes Association (ADA)
International Association of Diabetes and Pregnancy Study Group (IADPSG)
Major interests in research
1) Gestational diabetes mellitus
2) Preeclampsia3) Metabolism and nutrition of pregnant women and fetus4) Gynecological and Obstetrical Surgery

122

●Concurrent Symposium C-5.3

Uterine fibroids and IVF – What is the controversy?
Prof Zion Ben-Rafael
Tel Aviv, Israel
It is generally accepted that fibroids may cause infertility or pregnancy failure
depending on the number, size and location. Since our original study in 1995, manystudies have confirmed that in IVF settings, fibroids that distort the uterine cavity (submucous) may affectimplantation. Some of the studies also suggested that large intramural fibroids without submucous part cannegatively affect the results of IVF and hence should be surgically removed prior to IVF treatment. The
current dilemma mainly resides in the possible deleterious effect that intramural fibroids <7 cm, and even <5cm may have on implantation. The current results on the issue are conflicting which is probably related moreto the inability to properly evaluate the exact location of the fibroid and its imprint on the cavity.
The role of uterine fibroids as a cause of infertility remains controversial. The lack of randomized control
trials does not provide the means of establishing definitive causal relationships. The sharp decline inpregnancy rates in cases of submucous myomas appears to be quite convincing; however, the less significant
effect of intramural fibroids on the outcome is probably one of the reasons for the diverse results reported indifferent studies. New randomized studies that evaluate the size and location by 3-D ultrasound or MRI,
should be able to define more precisely which fibroids should be excised and when we can proceed with IVF,
despite the fibroids.
◆Biosketch
Prof. Zion Ben-Rafael,
Married + 3
Graduated from Tel Aviv University, Board certified in Israel and USA.
84-86, NIH Fellow in Reproductive Biology and Endocrinology, in Hospital of University of Pennsylvania,
Philadelphia.
Full Professor (1993), Associate Professor (1990), Tel Aviv University and Incumbent of the Tarnesby- Chairof Fertility regulation Tel Aviv University
Previously:
President, Israeli Menopause Association.
1997-2007 Chairmen, Department of OBGYN, Rabin Med Center.
1991-2007 Chairmen, Department of OBGYN, Golda-Hasharon.
1989-1991 Director, Katowitz Women's Health Center, Tel Aviv
Currently in charge of the IVF unit, Laniado hospitalFounder and Chairman, COGI - World Congress on Controversies in Obstetrics, Gynecology and Infertility.
1999 - Prague, 2001 - Paris, 2002 - Washington, 2003 - Berlin, 2004 - Las Vegas, 2004 - Bangkok, 2005 Athens. 2007, Barcelona 2007 - Shangahi, 2008 - Paris, 2009 - Beijin, 2010 - Berlin, 2011 - Paris, 2011 Hainan china.
Founder and Chairman of the Mediterranean Congress 1993-96
Founder and Co-Chairman, Gender based medicine, World Congress Berlin 2006
Co-Founder World congress on controversies in Obesity, Diabetes and Hypertension 2006
Scientific Publications:
Total over 350 Scientific Publications; 280 original articles, 2 books, editor 12 congress proceedings

123

●Concurrent Symposium C-5.4

A novel operative treatment of severe adenomyosis prior to embryotransfer
Hisao Osada
General Director, Shinjuku ART Clinic
Former Professor, Nihon University School of Medicine
Introduction: Severe case of adenomyosis is not only a cause of infertility, but may include severe
dysmenorrhea, thus may interfere with a woman's well-being. Routine conservative surgery for adenomyosis
involves a wedge resection of the involved uterine tissue, followed by approximating the remaining
myometrium and serosa. However, this method may retain unexcised affected tissue, and thus result in an
unsatisfactory post surgical prognosis such as being incapable of sustaining a normal pregnancy. Our
proposed treatment for severe cases of adenomyosis involves wide complete excision of affected tissues toreduce post surgical dysmenorrhea, followed by a triple-flap reconstruction of the uterine wall to prevent
ruptures in subsequent pregnancies.
Methods:
1. Resection and removal of all adenomyosis-affected myometrium: The affected tissue is vertically
incised, to split the area to be excised in two, the incision is extended to the uterine cavity. The tissue to be
excised is grasped and placed under tension with Martin forceps. The tissue is adequately dissected free withscissors, with care taken to retain a serosal flap with a layer of myometrium, as well as a medial flap
containing both endometrium and myometrium. The tissue flaps, both medial and distal must be more than 5
mm in thickness to assure adequate material for the reconstruction of the uterine wall. It is essential tointroduce an index finger into the uterine cavity to assure maintenance of an adequate medial flap thickness.
Special care must be taken to prevent damage to the Fallopian tubes.
2. Reconstruction of the uterine cavity: Care must be taken to retain sufficient endometrium to allow
reconstruction of an adequate uterine cavity. In cases of an over abundance of endometrial tissue, excess
amounts must be removed to secure a more physiological uterine cavity.
3. Reconstruction of the uterine wall: Reconstruction of the middle portion of the uterine wall involves
approximation of the myometrial musculature to ablate the space created by the excision of diseased tissue.
The serosa including adequate myometrium is dissected free with a scalpel to form the third flap. The serosal
or distal and third flap is then approximated to finish the reconstruction.
4. Hemostasis and application of hemostatic barriers for prevention of adhesion: The last step of thismethod is to apply TachoComb®, a Fibrin adhesive in sheet form, to the uterine surface for the control of
oozing. The applied TachoComb® is firmly anchored and works as physical barriers, thus contributing to the
reduction of post surgical adhesions.
Results: Clinical post surgical evaluation was performed using the Visual Analog Scale (VAS) to assess
dysmenorrheal and hypermenorrhea at 3, 6, 12, 24 months after surgery. We performed the procedures on 104
patients during the period between June 1998 and August 2008. Of the 26 women desired to conceive, 16(61.5%) subsequently conceived. Of these, 4 women conceived spontaneously and 12 women conceived by invitro fertilization and embryo transfer (IVF-ET). Two women who had IVF-ET experienced a spontaneousabortion; 14 went to term and all were delivered by elective Caesarean section. There were no cases of uterinecomplications to the pregnancies. The triple-flap reconstruction of the uterine wall following wide adequate
excision of adenomyosis tissue in women with hypermenorrhea and/or dysmenorrhea resulted in a dramatic124

●Concurrent Symposium C-5.4

reduction in both menstrual cramping and menstrual flow volume post surgically and gave women chances tobecome pregnant.

Severe Cases of Adenomyosis Surgically Treated by the Triple-flap Method

The mean (SD) VAS findings for menorrhagia were 3.27 (2.17) at 3 months,
2.89 (1.77) at 6 months, 2.63 (1.3) at 1 year, and 2.87 (1.77) at 2 year post-surgery◆Biosketch
Dr. Hisao Osada was born in 1946 and trained in the Department of Obstetrics and Gynaecology at Nihon
University School of Medicine in Tokyo, where he later served as Professor until 2009. He was at Mainz
University, Germany, from 1979 to 1981. His specialization includes laparoscopic surgery and reproductive
medicine. Dr. Osada currently teaches and practices at his university and at Shinjuku ART Clinic (General
Director) in Tokyo. He is the Vice-President of International Association of Private Assisted Reproductive
Technology Clinics and Laboratories, as well as the Vice Congress President of the 16th World Congress on
In Vitro Fertilization to be held in Tokyo in 2011.

125

●Concurrent Symposium C-5.5

The surgical management of uterine fibroids and adenomyosis
Michael DeRosa
Infertility Center of St. Louis
Uterine fibroids are present in approximately 70% of all women and 5 - 10% ofinfertile females. It has been found to be the sole abnormal finding in 1 - 2.4% ofinfertile women. Since childbearing is being delayed, couples are attempting pregnancy when female fertility
is in decline while the incidence of fibroids and adenomyosis are increasing. Numerous surgical procedures
such as hysteroscopy, laparotomy, laparoscopy, and robotics have evolved over the years giving us a myriadof choice. We will discuss the indications and methods of each of these procedures and the rational for when
they should be utilized for the treatment of uterine leiomyomas. We will also briefly discuss the surgicalapproach to adenomyosis.
◆Biosketch
Dr. Michael DeRosa, a graduate of St. Louis University School of Medicine and Residency program, has beenpracticing Obstetrics and Gynecology for nearly 25 yrs. His interest has always been directed towards thetreatment of the infertile patient and became affiliated with Dr. Sherman Silber and The Infertility Center of St.
Louis 17 yrs ago. Since that time he has been involved in IVF, ovarian tissue freezing and transplantation, andthe surgical management of the infertile patient.

126

●Concurrent Symposium C-6.1

The oncofertility: consequences of a previous oncological treatment andpossibilities
Rufino García-Otero Reina
Director, medico
The use of chemotherapy and radiation methods in patients of both sexes may haveirreversible consequences on fertility. Team work between the oncologist and the infertility specialist is
considered essential, especially in young patients who have not become mothers yet or those who
already have become mothers but still have a desire to increase their family.
The current use of methods such as the freezing of testicular or ovarian tissue, or vitrification of gametes has
improved the prognosis of such patients.
Oncofertility was born to provide a specialist, who masters both fields, and can devise pre or post cancer
treatment strategies to preserve fertility and who can integrate the work of oncologist and infertility
specialist together.
◆Biosketch
Specialist in Gynecology and Obstetrics by Landesaerztekammer Bayern Munich(Germany).
Third generation of gynecologists in the García-Otero family after his grandfather Dr.Rufino García-Otero
and his father, Dr. Eduardo García-Otero González. He studied in Germany, as well as his predecessors,
because the best and leading schools in Gynecology and Breast Pathology in the world are there.
5 years of work at the Klinikum Bayreuth. (Academic Hospital of the University of Erlangen-Nürnberg). Prof.
Dr. med. A. H. Tulusan (grade 1 reference centre in Germany for the treatment of gynecological and breast
cancer with a chemotherapy unit independent from the Frauenklinik, in addition to high risk perinatal
medicine).
During his stay at the Klinikum Bayreuth, participate in the following studies for patients with breast andgynecological carcinomas: ABCSG21, Compact, FACE, GAIN, Hera, BIS, NECTA, Pact, Panther, Preface,
Success.
Also, in the Klinikum Bayreuth , he manage the women's chemotherapy unit clinic for 6 months.
Since 2 years medical director of "Embryocenter CIVTE" (Clinic for IVF with 26 years experience) and
Senocur-Institute for Breast cancer". Both in Sevilla (Spain).
www.civte.com
www.senocur.com

127

●Concurrent Symposium C-6.2

Fertility Sparing in Stage I a2 to I b1-Cervix Carcinoma
Werner Lichtenegger
Charité Berlin
The incidence of pelvic lymph node metastases according to stage I a1 is very low. It
was shown by different publications from Bellin, Benedetti-Panici, Burghardt, under
3 mm invasion it lays by less than 1%, between 3-5 mm up to 3%. For the cervix important are the parametrial
nodes. Parametrial lymph node metastases without pelvic lymph node positivity are very rare. So, parametrial
involvement in patients with tumors less than 2 cm are under 1%. Rational for radical trachelectomy was thequality of life, the trend to more conservative surgery and the fact that in some cases the removal of theuterine fundus is not necessary. The question is: radical trachelectomy or only conization? When you look at
the literature radical vaginal trachelectomy shows positive nodes in about 4% and a lymph vascular spaceinvasion of about 29%. The recurrence rate in tumors less than 2 cm is very low, about 3%, over 2 cm itincreased up to 21%. In cases of radical vaginal trachelectomy pregnancy loss or delivery before 32 weeks
was in 44% of the patients. In cases of radical abdominal trachelectomy pregnancy loss or delivery before 32
weeks were 38% of the patients. Therefore we looked at the possibility of neoadjuvant chemotherapy
followed by conization in patients with I b1 and we found in these cases a neoadjuvant chemotherapy
pregnancy loss or delivery before 32 weeks only in 23% of patients. So, in conclusion fertility sparing in earlycervical cancer patients should be very well selected. Trachelectomies are an option in patients with I a2 but inpatients with negative nodes they are an overtreatment. In some series of trachelectomy up to 40% of thepatients did not have residual tumor after conization and trachelectomy results in about 40% pregnancy loss.
Neoadjuvant chemotherapy and conization had a pregnancy loss only for 23% and Paclitaxel/Carboplatin
weekly seems to be a very good and effective regimen.
◆Biosketch
Born on December 31st, 1942 in Graz/Austria
Since 1989 Chairman of the Gynecological Clinic of University Clinical Centre Rudolf Virchow in
Berlin/Germany
Since 1998 Director of the Clinic of Gynecology and Obstetrics of University Clinical Centre Charité in
Berlin/Germany, Campus Virchow Clinic
Research activities: surgical treatment and adjuvant therapy of cervical cancer, ovarian cancer andendometrial cancer; conducting of numerous monocentric and multicentric phase I, II and III studies;
identification of new molecular biologic prognosis factors in breast cancer, ovarian cancer and cervicalcancer; more than 300 publications and articles
Member of multiple international societies

128

●Concurrent Symposium C-6.3

Abdominal radical trachelectomy in our experience of 126 cases
Takuma Fujii, Hiroshi Nishio, Kazuhiro Minegishi, Naoaki Kuji,
Yasunori Yoshimura, Daisuke Aoki
Keio University, School of Medicine
In our institution, 140 abdominal radical trachelectomy (RT) were planned and 126
were performed from September 2002 through March 2011. The median age of the patients was 33 (23-44)
years. The majority of the lesions were stage Ia (21.4%) or Ib1 (78.6%). In histology, 87% (110/126) were
squamous and 13% (16/126) were adenocarcinomas. The median follow-up was 26 (2-104) months. Basedon the accumulated our experience in the past ten years, patients were offered radical trachelectomy if they
met the following criteria: 1.FIGO stage Ia with lymph-vascular space involvement or confluent invasion,
stage Ia2, stage Ib1 disease; 2. squamous carcinoma or well differentiated adenocarcinoma; 3.tumors less than
2cm in size or exophytic tumor less than 3cm in size but with little stromal invasion; 4. desire to preserve
fertility; 5. No evidence of disease outside the cervix. This criteria was reported by several authors, and we
also reported our experiences (Gynecol Oncol 115,51-55,2009). If the criteria above were satisfied in thecandidate patients, no recurrence was observed in two years. However, eligible criteria has not been
established yet. The risk of early invasive adenocarinoma was controversial because skip lesion of
adenocarcionma are present higher in the endocervical canal. Even if the surgical margin of the removed
cervix was negative, safety was not guaranteed. In our experience, 10% of patients planned RT wereabandoned and converted to hysterectomy due to positive lymph nodes or positive margins of the removed
cervix by frozen section in the original operation. Furthermore, additional 10% of patients who underwent RT
received adjuvant chemotherapy and/or radiation for those reasons. As a whole, 20% of candidate patients did
not meet the eligible criteria, consequently. There were some post operative complications; cervical stenosis,
amenorrhea and lymphocele. In order to prevent lymphocele, ligation of lymph vessels was critical andnon-suture technique for the retroperitoneum followed by Seprafilm (genzyme) treatment after
lymphadenectomy was also important. In the process of anastomosis of the neo-cervix to the vagina,
intrauterine device or cervicalplasty will improve to prevent cervical stenosis. In the obstetrical outcome, 11babies were delivered by Caesarean section. Eight of 11 patients conceived with artificial reproductive
technology. Nine babies were delivered after 32 weeks. Since no standard protocol for perinatal management
was established yet, perinatologists corresponded to individual pregnant women. RT seems to be an
oncologically safe procedure in well-selected patients with early-stage diseases. Obstetrical outcomes post RT
was tolerable. However, the patients need to be fully informed about perioperative and late complications
especially for the risk of premature delivery. Collaboration with gynecologic oncologists, perinatologists,
ART specialists and professional nurses were also critical issues for establishing this procedure.
◆Biosketch
1987 Graduated from Keio University, School of Medicine
1991 Research resident in Genetics Division of National Cancer Center Research Institute in Japan
1996 Research associate in Yale University, School of Medicine, Department of Internal Medicine, Medical
Oncology
1999 Gynecology, National Tochigi Hospital, Japan
2000 Assistant, Keio University, School of Medicine
2004 Assistant Professor, Keio University, School of Medicine

129

●Concurrent Symposium C-6.4

Laparoscopic radical trachelectomy
Masaaki Ando
Kurashiki Medical Center
Introduction: Vaginal trachelectomy with type II radical parametrectomy has been
well reported and although rare, recurrences were reported at the lateral parametrium.
Our concerns with the inadequate resection of the parametrium and operability in narrow vaginal access cases
led us to develop a Type III total laparoscopic counterpart focusing on the isolation and removal of the total
length of the parametria. Although there are some concerns that a Type III radicality may have a negative
impact on fertility, to date we are happy to report a total of 9 births.
Methods: From 2001 to 2010, 50 patients with stage 1a2 to stage 2a1 cervical cancer underwent our total
laparoscopic radical trachelectomy. The inclusion criteria was exophytic type with a size of less than 2.5cms
with at least 5mm residual cervix with a surgical clearance of at least 1cm. Squamous cell carcinoma andadenocarcinoma cases were included. Cases with lymph node metastasis discovered after MRI or CT were
excluded.
To begin the procedure, first we create a vaginal cuff in order to define the exact length of the vaginal andprevent spillage of the tumor into the abdominal cavity. We begin the pelvic lymphadenectomy using ourumbilical ligament suspension technique. After isolation and preservation of the uterine artery, and complete
isolation of the ureter the pararectal space is developed isolating the cardinal ligament. The cardinal ligamentis then transected at the pelvic sidewall in a Type III radical fashion using a vessel sealing system or linear
stapler. All other paracervical ligaments as well as the upper 1.5cms of the vagina are transected. Then thecervix was cut from the uterine body at the level of 5mm below the internal os. The sump of the vaginal wasanastomosed to the residual cervix laparoscopically.
Results: Average duration of the procedure was 393±109min and blood loss measured 447±330mL. No
complications occurred except for temporary lymphedema. Although one case with lymphatic space invasion
died from recurrences at multiple sites, the other 24 cases are alive with no evidence of disease after 6 to 72
months.
At the time of the operation 33 patients were single with no plans for pregnancy. Only 12 cases havereportedly attempted pregnancy, of these 7 cases have reportedly become pregnant one time, 3 cases havebecome pregnant a second time. All but 2 cases have become pregnant as a result of ART or other fertility
treatment. After pregnancy, three cases experienced blighted ovum. One case, terminated at 21 weeks in her
first pregnancy terminated again at 18 weeks in her second pregnancy both times due to PROM andintrauterine infection. Other pregnancies have resulted in 9 live births all delivered by cesarian section. One
patient delivered twins (MCT) at 23 weeks due to PROM. Although one twin died before delivery the othersurvived and is healthy at time of writing. Two cases delivered at 26 weeks, one case at 28 weeks and one
case at 31weeks, one case at 34weeks one case at 36weeks. Two case was delivered at term.
One baby died due to congenial heart disease. Two cases underwent Sailing technique to prevent intrauterine
infection. One procedure deemed effective as the case achieved term delivery. Another procedure failed andterminated at 23 weeks (twin pregnancy case).
At time of writing there are 2 other on going pregnancies. One is 23 weeks while another is 29 weeks. It has
been impossible to follow up on all cases as some were referred from distant places and have not reported
their condition.
Discussion: Type III radical transection of the cardinal ligament is possible totally laparoscopically preserving
the uterine artery so even cases suffering from malignant disease can benefit from new procedures that
improve the quality of life. In spite of the radical nature of the procedure compared with the radical vaginal130

●Concurrent Symposium C-6.4

trachelectomy, pregnancy is possible. A recent paper related to vaginal radical trachelectomy reported a 4%
recurrence rate with half of those recurrences happening in the pelvic sidewall. To date we have experienced
no recurrent cases meaning that the oncologic requirements have been fulfilled while preserving the patients‘
fertility potential.
◆Biosketch
1973-1980 Jichi Medical School
1980.04
Okayama Red Cross Hospital: Obstetrician
1984.04
Kurashiki Medical Center: Head of Gynecology Department.
2001.04
Osaka University: Visiting Lecturer of Obstetrics and Gynecology.
2003.04
Kyoto University: Visiting Lecturer of Obstetrics and Gynecology.
2004.08
Beijing Capital University: Visiting Professor of Obstetrics and Gynecology.
2007.04
International Advisory Board: Society for Laparoscopic Surgeons (USA)2009.03
Thai-German Multidisciplinary Endoscopic Training Center Visiting Professor of Obstetrics andGynecology
2006.03
Keio University School of Medicine: Visiting Associate Professor of Obstetrics and Gynecology
2009.03
Kurashiki Medical Center: Head of Center for Endoscopic Surgery
2009.03
Kurashiki Medical Center: Vice Director
2009.03
Kurashiki Medical Center: Trustee
2009.04
Keio University School of Medicine: Visiting Professor of Obstetrics and Gynecology
2010.04
Clinical Professor Kyoto University School of Medicine: Clinical Professor Obstetrics andGynecology
2010.04
Clinical Professor Osaka University School of Medicine: Clinical Professor Obstetrics andGynecology
2011.04
Honorary member of Austlarian Gynecologic Endoscopic Society131

●Concurrent Symposium C-7.1

Cryopreservation of human oocytes of cancer patients in Japan
Shokichi Teramoto
Shinbashi Yume Clinic
Given the established egg-freezing technology, institutions in Japan affiliatedwith the International Association of Private Assisted Reproductive Technology
Clinics and Laboratories are currently engaged in clinical research projects to collect oocytes from women
who may become infertile due to treatment of hematological malignancies, such as leukemia, before or during
treatment. The oocytes will be cryopreserved so that the affected women can still become pregnant in the
future. This activity is based on the belief that the preservation of fertility in women who wish to give birthwill contribute to a better quality of life once they have overcome the underlying disease.
The most important thing to consider when collecting oocytes from patients with hematological malignancy
is not to interfere with the treatment protocol for the underlying disease. To ensure this, an ovarian stimulation
technique that is compatible with the cycle of the treatment for the underlying disease and has flexible startand end dates for ovarian stimulation is required. In addition, since antitumor therapy is usually administeredrepeatedly after each hematological recovery, the egg collection technique also needs to be repeatable. For
these reasons, the maximum stress that can be imposed by egg collection on the ovary and the patient's bodyare strictly limited, making the conventional long protocol the least appropriate strategy for ongoinghematological malignancies with poor hematological status, except for very early-stage malignant lymphoma.
Both the short protocol;SP and antagonist protocol;AP are less stressful than the LP, but are not repeatable
and less flexible in terms of start and end timing. On the other hand, the clomiphene protocol;CP is repeatableand flexible. The letrozole protocol;LP and the natural cycle protocol;NP are highly comparable in terms of
repeatability and flexibility and also the least stressful, but are associated with a relatively high probability ofcancellation. Thus, the SP and AP can be used for low-grade malignant lymphoma for which the initiation ofremission induction can be delayed, whereas the CP should be chosen for hematological malignancies forwhich remission induction has been successfully completed and treatment is proceeding to consolidation
therapy. The LP and NP are applicable to all stages of disease as they are associated with relatively low stresson the ovary and the patient's body. They also carry a low risk for complications, such as ovarian hemorrhage,although efforts are required to reduce cancellation and increase the number of oocytes that can be collected.
When collecting oocytes, safety considerations are required for anesthesia and prevention of infection andbleeding. Anesthesia is not required when collecting one or two oocytes, but is required when the procedure is
expected to take a long time and when multiple punctures are required. General anesthesia may cause a delay
in implementing urgent measures, such as blood transfusion, as it may mask signs of ovarian hemorrhage e.g.,
abdominal pain. Therefore, pain management should be done with local anesthesia, which does not reduce
consciousness and enables information indicating abnormality, such as abdominal pain, in to be obtained in atimely manner.
Since 2007, we have succeeded in effectively removing pain by injecting a small amount of lidocainehydrochloride into a gap between the vaginal muscularis and peritoneum under ultrasound guidance. Infection
can be completely prevented by vaginal disinfection using povidone iodine and eliminating the process offollicle washing. This requires a technical basis to avoid the entrance of povidone iodine into the collection
solution and ensure reliable egg collection with only one aspiration procedure. To prevent ovarian hemorrhage,
it is essential to introduce an egg collection technique that can reduce the number of punctures into the ovarianparenchyma and a fine egg collection needle. We have developed unique fine egg collection needles since1998, and while constantly improving these tools, we have used them for collecting oocytes from leukemia

132

●Concurrent Symposium C-7.1

patients since 2001. We have also used 22- and 23-gauge tapered needles in practice since 2006 and havesucceeded in collecting oocytes with a very high level of safety.
The best practice is required for egg collection from patients with hematological malignancy in all fields ofassisted reproduction technology, including freezing, ovarian stimulation and egg collection techniques. The
following presentation describes our efforts and achievements, which have been made over a period of morethan 10 years.
◆Biosketch
Born in 1957 in Japan, Shokichi Teramoto graduated in 1990 from Kanazawa University School of Medical
Science, Ishikawa, Japan. Subsequently he trained in the Department of Obstetrics and Gynecology at theGraduate School of the same university. From 1994 he trained as an infertility specialist under the guidance of
Dr Osamu Kato, in 1996 becoming Vice Director of Kato Ladies Clinic, an IVF center in Tokyo. Since then,he has devoted himself to establishing an ovulation stimulation method using clomiphene citrate. Other
interests include obtaining oocytes from leukemia patients and building an oocyte retrieval-cryopreservation
system for leukemia patients.

133

●Concurrent Symposium C-7.2

Cryopreservation of human ovarian tissue, clinical data and scientificconsiderations
Claus Yding Andersen, MSc, DMSc, Professor,
Laboratory of Reproductive Biology, Section 5712,
University Hospital of Copenhagen, 2100 Copenhagen, Denmark
Girls and women suffering from a cancer or another disease that require treatment
with gonadotoxic drugs may as a side effect reduce the ovarian pool of follicles. The gonadotoxic effect isdependant on the specific treatment and is influenced by the dose of the therapeutical agent and the possible
use of radiotherapy. Especially, alkylating substances and radiation to the abdomen in case of cancer diseases
often cause irreversible damage to the ovaries. When the ovaries are depleted of follicles many womenexperience profound effects on the physical and psychological status. Menstrual cycles ceases and it is notpossible to become pregnant. To young girls it may further mean that normal puberty fails.
Cryopreservation of ovarian tissue is a new method, which has been developed in an attempt to circumventthe long-term ablative effect on reproductive performance by gonadotoxic treatment. Removing one wholeovary or part of an ovary from women in their reproductive years prior to treatment and cryopreserving thetissue can keep a viable pool of follicles. When the women have been cured and is considered fit, the thawed
ovarian tissue may be transplanted to those who entered menopause.
In Denmark cryopreservation of ovarian tissue has been organized with one central laboratory that freezes all
the tissue in close collaboration with three fertility clinics round the country. Totally more than 500 girls andwomen have had ovarian tissue cryopreserved in Denmark. The youngest girl was 0,5 years old and the oldest
38 years. We have currently cryopreserved ovarian tissue from around 100 girls younger than 18 years of age.
The ovarian tissue is excised at the local hospital and transported on ice to the freezing facility, where
cryopreservation and storage is performed. In case of transplantation the frozen tissue will transported to the
local hospital for the operation. This transport model has been validated and has now been used for more than
250 cases.
In Denmark a total of 18 women have experienced transplantation of frozen/thawed ovarian tissue a total of
25 times (7 women having tissue transplanted twice). All women regained ovarian function and none haveexperienced relapse as a consequence of the transplantation. Over a period of 20-25 weeks levels of FSH
gradually return to pre-menopausal levels and menstrual cycles are regained. The longevity of the tissue
depends on the age of the woman at tissue retrieval and the amount of tissue transplanted. Most womenexperience return of ovarian function for some years with just a fraction of tissue from one ovary being
replaced. Recently, one child has had ovarian tissue transplanted for natural induction of puberty; this casewill be presented in detail.
Six women have been pregnant; in most cases following natural conception. Two women have delivered threehealthy babies as a result of transplanted frozen/thawed ovarian tissue. In the latter two cases the tissue wastransported 4-5 hours prior cryopreservation. The presentation will review our experiences and results withtransplantation of cryopreserved ovarian tissue.
◆Biosketch
Claus Yding Andersen is professor in human reproductive physiology at the Faculty of Health Sciences,
University of Copenhagen, Denmark and has since 2009 been heading Laboratory of Reproductive Biology at
the University Hospital of Copenhagen, Denmark. Claus Yding Andersen qualified first as MSc from theDanish Technical University (1979) and obtained his Doctor of Medical Science degree from University of
Copenhagen in 1997. His main research interests include cryopreservation of gonadal tissue, ovarian
endocrinology and human embryonic stem cells. He has published more 200 scientific papers and has given alarge number of international presentations.

134

●Concurrent Symposium C-7.3

Conservative Treatment of Borderline Ovarian Cancer
A.H.Tulusan1; V.Russu1; B.Lex1; M.Popovic1; I.Sopov1; H.Volkholz2
Dept. Gynecology and Obstetrics1 and Dept. Pathology2; Klinikum
Bayreuth, Germany
Borderline ovarian Carcinoma (BOC) are epithelial neoplasm with pathological characteristic in between
benign and frank malignant ovarian cancer. So the WHO define the BOC as a Ovarian Tumor with low
malignant potential (LMP). Despite peculiar histopathological features they generally are associated with anexcellent prognosis. Most series reported a 5-years survival rates of 100% for LMP stage I or IIA. Even for
tumors involving the pelvis or abdomen still the 5-years survival rates are about 80%. Still much effort has
been and to be done to identify the features associated with poor prognosis of some of the LMP.
Beside architectural complexity with papillary and detached atypical cell clusters also nuclear atypia, mitotic
activity and lack of invasion are used for the diagnosis of LMP. Even then there are still often difficulties todistinguish between LMP and frank invasive ovarian cancer especially in cases with dissemination andimplantation of LMP tumor cells seeds at the peritoneal cavity.
Histological different types of LMP was distinguished: serous, mucinous, endometroid, clear celland they
could have different prognosis.
Patients with LMP are generally younger (45Y) than patients with invasive ovarian cancer (56Y).
Only 30% of the LMP (stage I) are growing bilaterally, higher stage cases are bilateral in 60%.
Only 25% of stage I LMP have elevated CA125.
Even treated with complete hysterectomy and bilateral adnexectomy some LMP has a late recurrence (16years) some with a fatal outcome. Some investigator showed a poor prognosis for cases with high MIB-1 or
Ki-67 on cases with late recurrence.
Surgery is still the corner stone of Ovarian Cancer management. This is also true for LMP. Even without
adjuvant chemotherapy the survival rates of LMP ranged from 90%-100%. To have the better possibility of
distinguishing prognosis of LMP the same detailed staging like for invasive ovarian cancer has to be done.
More than 80% of the LMP present with stage I disease.
Since many LMP patients are in the childbearing years the possibility of conservative treatment must beseriously considered.
No evidence of adverse effect of conservative fertility preserving surgery for stage I LMP in terms of survival
has been seen. But conservative treated LMP patients can have a higher recurrence rate. The need of adjuvant
systemic therapy for higher stages of LMP is still a subject of debate. Randomised studies on LMP stage I
showed no necessity for adjuvant treatment (Creasman et al.1982; Young et al. 1990).
For more advance stages of LMP (II – IV) the surgical therapy of LMP is similar to patients with invasive
ovarian cancer. Patients with aneuploid tumors have a worse prognosis compared with diploid tumors (Tropeet al, 1990). In these cases Platin containing chemotherapy are indicated.
Experience with the conservative treatment of 5 cases of LMP in our institution will be presented.
Conclusions: For the individualised risk adapted treatment of LMP ovarian tumors and considering thepreservation of fertility following must be done:
1. Interpreting the stage and histological and biological risk factors.
2. Determine surgical management and the possibility and need for fertility preservation.
3. Determine the appropriate medical chemotherapy management for advanced disease.
◆Biosketch

135

●Concurrent Symposium C-7.3

Agustinus Harjanto TULUSAN, M.D., Ph.D., German, born on 21 July 1945
Present Position:
Director, Dep. of Gynecology and Obsterics Bayreuth, Academic Teaching Hospital of the Erlangen
University, Klinikum Bayreuth, Preuschwitzer Str. 101, 95445 Bayreuth, Germany
Professional Qualification:
Medical Education: University Erlangen 1966-1972
Habilitation/Ph.D 1982 University Erlangen: ―Carcinoma lobular in situ and breast cancer‖
Dept. of Pathology: 1972-1976 University ErlangenDept. Gynecology and Obstetrics: 1976-1994 University ErlangenDept. Gynecology and Obstetrics: since 1994 Klinikum Bayreuth
Previous appointments:
Professor of Gynecologic Oncology and Head of the Dept. of Gynecologic Onkology
Womens Hospital University Erlangen: 1985-1994
Vice Director of the Dept. of Gynecology and Obstetrics, Univ.Erlangen: 1986-1994
Membership of medical scientific societies:
1. German Gynecologic and Obstetrics Society (DGGG)
2. German Senology Society
3. Bavarian Gynecology and Obstetrics Society
4. German Cancer Society (Deutsche Krebsgesellschaft)
5. Gynecology Oncology Study Group (Arbeitsgemeinschaft Gyn. Onkologie)
6. Reconstructive Surgery Study Group (Arbeitsg. Wiederherstellende Operation)
7. Gynecologic Pathology Study Group (Arbeitsg. Gyn. Pathologie)
8. Society Pelvic Surgeons since 2003
9. Member of the American Society of Clinical Oncology since 2007136

●Concurrent Symposium C-7.4

Infertility, ovulation induction treatments and the incidence of breast,
ovarian and endometrial cancers – Thirty years of follow up
Lunenfeld1 B and Gertner L2
Faculty of Life Sciences, Bar Ilan University, Ramat Gan, Israel1
Women and Children's Health Research Unit, Gertner Institute for
Epidemiology and Health Policy Research, Tel Hashomer, Israel2
Objective:
To evaluate the possible risk for cancer development in infertile women treated with ovulation inducing
agents.
Design: The study is a historical cohort analysis of 2431 women who were treated for infertility at the Sheba
Medical Center during the period 1964-1974 and contributed more than 84,000 women years of follow-up.
Material & Methods: Cancer development was assessed through linkage with the National Cancer Registry
updated to 31.12.2005. Standardized incidence ratios (SIR) were calculated between the observed cancer
cases and the expected cancer rates in the general population with respect to age and continent of birth.
Birthrate was assessed through linkage with the National Birth registry.
Results: The cohort includes 33.8 years of follow-up with a mean age at the end of follow-up of 62.7 years.
Over all 350 cases of all-site cancer were observed as compared to 338.4 expected (SIR=1.03; 95%CI
0.93-1.15). Eighteen cases of ovarian cancer were observed as compared to 18.1 expected (SIR=1.0; 95%CI
0.59-1.57). For breast cancer, 153 cases were observed as compared to 131.9 expected (SIR=1.16; 95%CI
0.98-1.36) and for endometrial cancer 30 cases were observed as compared to 17.8 expected cases (SIR=1.69;
95%CI 1.14-2.41).
Conclusion: Hormonal causes of Infertility were found to be associated with increased risk of endometrial
cancer. Borderline increase in the risk for breast cancer was observed in infertile patients. Ovarian cancer risk
was not found to be elevated. No significant changes in the risk were found following treatment withgonadotropins◆Biosketch(For Professor Lunenfeld‘s Biosketch, please refer to his abstract for Concurrent Symposium C-3.1.)

137

●Concurrent Symposium C-8.1

Highly efficient and safety vitrification using hydroxypropyl cellulose as a
macromolecular supplement for cryopreservation of oocytes andblastocysts
Luis Ruvalcaba, Martha Isolina Garcia Amador, Rocio Martinez Armas
Instituto Mexicano de Infertilidad
Introduction: Vitrification as an alternative method of cryopreservation on human assisted reproductive
technologies (ART), resulting significantly improved survival rates of oocytes and embryos. Reduce risk ofviral contamination resulting from use of media containing biological macromolecules, government regulators
in some countries have stipulated that non-serum substitutes must be used in human ART, including
cryopreservation. Study was to test efficacy of a synthetic macromolecule, hydroxypropyl cellulose (HPC), in
vitrification.
Methods: Four experiments were conducted using bovine oocytes, blastocysts, and human oocytes, 4-cellembryos. Bovine oocytes retrieved from ovaries harvested at slaughter at an abattoir were subjected to in
vitro maturation (IVM), in vitro fertilization (IVF), in vitro culture (IVC). Used either as Metaphase II oocytes,
blastocysts. In each of two experiments, three groups of bovine oocytes (Experiment 1) and bovine
blastocysts (Experiment 2) were cryopreserved with Cryotop method. Group 1, the vitrification medium with
0.6 mg/ml HPC, and Group 2, with 1% commercial Synthetic Serum Substitute; Group 3 contained no added
macromolecule. Experiments were repeated twice with total of 90 oocytes, 90 bovine blastocysts were used.
(Experiment 3), human oocyte were vitrified and medium containing 0.6% mg/ml of HPC. This experiment
was repeated several times for total of 30 oocytes had been vitrified. After vitrification, survival was basedon development into blastocysts of oocytes subjected to IVF and IVC. Blastocysts were classified as
survivors if they reformed a blastocoelic cavity after 3 hours in culture. (Experiment 4) 35 human oocytefrom 9 patient and 7 blastocysts from 4 patient were cryopreserved with ES, VS, TS, DS, WS containing 0.6
mg/ml HPC. We excluded patient with PCO, Low response, patient over 35 years old or implantation
failure.
Results: (Experiment 1), The respective morphological survival rates of oocytes Groups 1 (100%), 2 (100%)and 3 (87%). After fertilization and culture, the corresponding rates of two-cell formation in these threegroups were 63%, 67% and 37%. The respective rates of blastocyst formation were 20%, 17%, and 7%.
(Experiment 2), survival rates of bovine blastocysts of the three groups were 100%, 100% and 93%,
respectively. (Experiment 3), all 30 human oocyte vitrified in HPC exhibited normal osmotic responses
during removal of the cryoprotectants. (Experiment 4) survival rates were 100% on oocytes and on blastocysts
all 42.
Conclusion: High morphological and functional survival of bovine oocytes vitrified in medium supplemented
with HPC, as well as morphological survival of bovine blastocysts and human oocyte suggest that (HPC) may
be a safe and effective supplement for vitrification in human ART.
◆Biosketch
Luis Arturo Ruvalcaba Castellón M.D. Graduated from University of Autonoma de Guadalajara, Mexico.
Director of the Mexican Institute of Infertility in Guadalajara, MexicoChief of the Department of Gynecology
and Obstetrics at the Medical Center Specialty Hospital of Puerta de Hierro in Guadalajara, Mexico.
Laparoscopic Surgery Fellow at the University of Kiel, Germany, 1994 Fellow Biology of Reproduction,
Assisted Reproduction at the Central Hospital Baptist, Lexington, Kentucky, USA and North Carolina Chapel
Hill, 1997. Fellow Assisted Reproduction and Vitrification in Kato Ladies Clinic, Tokyo, Japan 2004. Active
member of International Society for Preservation Fertilty, ESHR, AMMR, APART. Robotic Surgery Da Vinci,
Florida 2010. Pioneer in Vitrification in Latino America.

138

●Concurrent Symposium C-8.2

Role and Possibility of Vitrification in Human Oocyte
Masashige Kuwayama
Repro-Support Medical Research Center, Tokyo, Japan
Recent drastic advances in cryobiology have made it possible to preserve various
types of reproductive cells with little viability loss. Ultra rapid vitrification, thealternative cryopreservation method seems to be a powerful tool to any biological specimens, which cannot bepreserved by the conventional slow freezing and vitrification. Ultra rapid vitrification realized the successful
clinical use of vitrification not only for human PN zygotes, cleavage stage embryos and blastocysts but also
for oocytes. Cryopreservation of oocyte means to give a time between the day of aspiration of oocytes and
IVF for ET. This is called Oocytes Bank. Oocyte bank keeps the possibility to have a baby in the future family
life for all the women who have any risks of future infertility. The women of blood cancer, reproductive organcancer and breast cancer have risks of losing ovarian function by a side effect of the treatments. Turners
syndrome girls will lose ovarian function in their early 20s. Ovarian function can be also lost by a case of
accidental exposure to radiation by a nuclear power station. Oocytes bank gives patients not only a time but
also a distance for IVF treatment. Oocytes bank eliminates the difficulties to synchronize the cycles between
the donor and recipient in the egg donation IVF programs over national borders. Sex reassignment surgery has
been recently legally approved in some countries including Japan. Some FTM women want to have babies
with her own gene in their future family life after the surgery and marriage with women. In addition, allwomen actually lose their ovarian function in their advanced age by senescence of the body. Self oocyte
banking has been already used for career women for the happy family life with own children for the delayeddelivery over 40 years old, as an option of the personal fertility plan in many countries. Ultra rapid
vitrification, the Cryotop method has been clinically applied to more than 500,000 cases (30,000 cases for
oocytes) for these 10 years in 40 countries with excellent clinical results without any problems like virus
contamination during storage. Safety and effective technology of oocytes cryopreservation has been alreadyestablished, and women have now choices of the solutions for the loss of ovarian function toward having theirown babies to improve quality of life as women.
◆Biosketch
Name: Masashige Kuwayama Repro-Support Medical Research Center
Ph.D.: Hokkaido University
Occupation:
Present- CEO, Repro-Support MRC; 2010- General Academic Supervisor, Kato Ladies group; 2007- Board
Internal Society of MAAR; 2005- ditor, J. Reproductive Bio Medicine Online; 2005- Board, JSClE; 2005Lecturer, IUMW, Meiji, Azabu Univ.
Publications: 81 Papers on Scientific Journal: JRF, Reproduction, Cryobiology, CryoLetters, BOR etc.
290 Papers on Scientific Meeting: IETS, ICAR, ASRM, ESHRE etc.
Topics: Scientific Director, largest IVF center for 10 years.
R & D Director: vitrification, GVT, spermatogenesis, ES-cells.
Obtained first calf after vitrification of blastocyst (1991), MII oocyte (1992). First vitrification porcine
pre-hatch blastocyst, oocyte (1997, 1998). Developed Cryotop method for human oocyte (1999), embryo(2000), first oocytes bank (2001), in USA (2003). Vitrified 500,000 oocytes and embryos in 40 countries by
Cryotop method.

139

●Concurrent Symposium C-8.3

Blastulation and pregnancy rates after vitrified human zygote culture
for 4 days: Preliminary results
Safaa Al-Hasani, Beate Schöpper, Ehab Abu Marar, Tim Cordes,
Askan Schultze-Mosgau, Klaus Diedrich and Georg Griesinger
Women Hospital, University of Schleswig-Holstein, Campus Luebeck,
Ratzeburger Allee 160, 23538 Luebeck-Germany
Introduction: Blastocyst culture has been introduced with the aim of increasing the efficacy of embryo
selection. In this preliminary study, blastocyst culture was offered to patients, who had shown excessive
ovarian response and thus had all 2 PN stage oocyte frozen by vitrification in order to prevent ovarian
hyperstimulation syndrome (OHSS).
Material and Methods: 29 patients were included in this study till now. These patients were stimulated witheither corifollitropin alfa or recombinant FSH in a GnRH-antagonist protocol. Final oocyte maturation was
triggered with GnRH agonist to avoid OHSS. The fresh embryo transfer was cancelled and all 2PN stage
oocytes were vitrified by the Cryotop method (Al-Hasani et al. (2007). Four to six zygotes were warmed per
attempt to transfer and cultured in ―Sage sequential media‖ for further 4 days under oil. A maxiumum of two
blasotcysts were transferred in a programmed cycle and if more blastocysts were available they werere-vitrified.
Results: A total of 160 zygotes were warmed from 29 patients till now and 42 embryos reached the early andexpanding blastocyst stage, while 9 reached the morula stage (32%).
The scoring system used was according to Gardner et al. (1999). The implantation rate achieved in this study
was 31.4 % and the pregnancy rate was 34.5%.
Conclusion: These results show that blastulation rate after vitrification is high, and thus can be offered topatients with a sufficiently high number of 2 PN stage oocytes. In combination with agonist triggering, OHSS
can be avoided while efficacy is high.
References:
Al-Hasani S, Batuhan Ozmen, Nikoleta Koutlaki, Beate Schoepper, Klaus Diedrich, Askan Schultze-Mosgau(2007): Three years of routine vitrification of human zygotes: is it still fair to advocate slow-rate freezing?
RBM online 14, 288-293
Gardner DK, Schoolcraft WB. In vitro culture of human blastocyst. In: Jansen R, Motimer D, eds. Towards
reproductive certainty: infertility and genetics beyond 1999. Carnforth: Parthenon Press, 1999: 378-88.
◆Biosketch:
Dr Safaa Al-Hasani graduated from the University of Baghdad in 1977. In 1980 he undertook his doctorate
degree, as well his specialty, in the field of reproductive veterinary medicine from Tieraeztliche Hochule inHannover. He has been the director of assisted reproduction laboratories in Erlangen, Bonn and Lübeck sincethat time. In 1982, he was in the scientific group that achieved the first German IVF baby in Erlangen.
Thereafter, he became a staff member of the University of Lübeck where he became an extra-ordinary
Professor in 2002 and where he is still working. He is the author or co-author of more than 250 scientificpublications.

140

●Concurrent Symposium C-8.4

Cryopreservation in human Assisted Reproductive Technology
Stanley P. Leibo
Department of Biological Sciences, University of New Orleans,
New Orleans, LA, U.S.A.
AIMS: The purpose of this lecture is to briefly review applications ofcryopreservation to the practice of human Assisted Reproductive Technology(ART).
DESCRIPTION OF RESULTS: In 1953, the first human pregnancies resulting from artificial insemination withfrozen-thawed spermatozoa were reported. Exactly thirty years later, the first human pregnancy resulting fromtransfer of a frozen-thawed embryo was reported, and the first birth from a cryopreserved embryo wasreported one year later. Since then, cryopreservation of reproductive cells and tissues has become an essential
technique in the ART clinical laboratory. Babies have been born from women impregnated with semen thathas been stored at -196°C for more than 25 years. As for embryos, in 2005 in Canada alone 464 babies wereborn from frozen-thawed embryos; in that same year in Europe more than 13,000 pregnancies were producedby transfer of frozen-thawed embryos. From 2005 to 2009 in the United States more than 37,000 pregnancies
resulted from cryopreserved embryos. Although difficult to document precise figures, it can be roughly
estimated that many more than 1,000 babies have now been born after fertilization of oocytes cryopreservedby any one of several techniques. More recently, pregnancies and births have resulted from various methods
to utilize reproductive tissues, such as ovaries and testes, that have been cryopreserved.
CONCLUSIONS: Thirty-three years after the remarkable report by Steptoe and Edwards of the birth of the firsthuman child by assisted reproduction, millions of other children have been born by application of similar
methods. Cryopreservation of reproductive cells and tissues has contributed to the efficiency and success ofhuman ART.
◆Biosketch
Stanley P. Leibo received his Bachelor of Arts degree from Brown University, Master of Science degree fromthe University of Vermont, and Master of Arts and Doctor of Philosophy degree in Biology from Princeton
University. He began his career in cryobiology with Peter Mazur at Oak Ridge National Laboratory in
Tennessee. He then became Vice-President of Research and Development at Rio Vista International, a cattlecompany in San Antonio, Texas. In 1988, he was appointed Associate Professor of Obstetrics and Gynecology
and of Urology at Baylor College of Medicine in Houston. From 1991 to 1998, Dr. Leibo was Professor of
Biomedical Sciences at the Ontario Veterinary College of the University of Guelph in Ontario, Canada. In
1998, he was named to his present positions as Professor of Biological Sciences at the University of New
Orleans, and Senior Scientist at the Audubon Center for Research of Endangered Species in New Orleans.
In addition to his permanent positions, Dr. Leibo has served as a Fellow of the Japan Society for the
Promotion of Science at Kyoto University, and for four years was Visiting Professor on the medical faculty at
the Dutch-speaking Free University in Brussels, Belgium. He was elected president both of the Society for
Cryobiology and of the International Embryo Transfer Society, and was named an Honorary Lifetime Memberof the American Embryo Transfer Association. In 2009, Leibo received the Pioneer Award of the International
Embryo Transfer Society to recognize his role in the derivation of the original methods to cryopreserve
gametes and embryos. He has published more than 135 scientific articles and book chapters, and has delivered
more than 180 lectures in the United States and in twenty-nine other countries.

141●Concurrent Symposium C-9.1

Evaluate the IVF Success Rates of Accumulated Pregnancy Rates per
Oocyte Retrieval
WU Chun-Xiang MD, LIU Jia-Yin MD, PhD.
The Center for Clinical Reproductive Medicine, The First AffiliatedHospital of Nanjing Medical University, Nanjing 210029, China, 210029
Objective: To assess the efficiency of oocyte utilization after controlled ovarian
stimulation.
Methods: We estimated cumulative pregnancy rate per oocyte retrieval among patients undergoing IVF/ICSI
between 2009 and 2010 at the Center for Reproductive Medicine, Nanjing Medical University. Couples were
followed-up until either discontinuation for no embryo to transfer or pregnancy with visualization of a fetal
heartbeat on ultrasound. Analyses were stratified according to maternal age.
Results: A total of 1,593 oocyte retrievals yielded 16,118 oocytes. Mean age (±SD) was 30.3 (±4.3) years,
mean number of transfer per oocyte pickup was 1.2, mean number of embryo transferred was 1.7. The
cumulative pregnancy rate per oocyte retrieval was 70.2%, the multiple pregnancy rate was 27.9% and themiscarriage rate was 8.6%. While the same number of embryos transferred, no statistically significant
difference was detected between fresh and the FET cycles with respect to pregnancy rate, implantation rate,
multiple pregnancy rate and miscarriage rate. Among patients who were older than 35 years of age, thecumulative pregnancy rate was significant decreased (P<0.05), and the miscarriage rate was significant
increased (P<0.05).
Conclusion: During ART cycles, maternal age remain a factor that influence the cumulative pregnancy rate
per oocyte retrieval, reducing the number of embryos transferred does not influence the final success rate,
while the multiple pregnancy rate was significant lower, this strategy yields a lower pregnancy in a fresh IVF
cycle, but the difference is almost completely overcome by an additional frozen embryo transfer cycle.
◆Biosketch
JIAYIN LIU, M.D., Ph. D.
Full professor of Obstetrics and Gynecology,
Director of The Center for Reproductive Medicine,
Department of Obstetrics and Gynecology,
Jiangsu Province Hospital.
Chief doctor on Reproductive Endocrinology, The First Affiliated Hospital of Nanjing Medical University.
Vice director of Jiangsu Province Women‘s and Children‘s Health Center,
Board member of Chinese Society of Reproductive Medicine.
Board member of Chinese Society of Family Planning.
Vice- director of Group on Gynecologic Endocrinology in Chinese Society of Obstetrics and Gynecology,
Vice- chair of Jiangsu Society of Obstetrics and Gynecology,
Vice- chair of Jiangsu Society of Genetic Medicine.
Vice-chair of Jiangsu Society of Women‘s Health Care
President of Jiangsu Association of Integrative Medicine on Reproductive Medicine

142

●Concurrent Symposium C-9.2

Genetics and Health of Children Born from Cryopreserved Oocytes
Ilan Tur-Kaspa, MD.
Institute for Human Reproduction (IHR) and the Department of Obstetrics
and Gynecology, The University of Chicago, Chicago, IL, USA.
Introduction: The safety of new technologies or treatments in ART is of criticalimportance to patients and practitioners. Oocyte cryopreservation has major benefits in modern ART: Fertility
preservation; circumvention of the ethical, legal and religious implications associated with embryo storage;
optimization of egg donation programs; and for embryonic stem cell research. We investigated the safety of
the clinical use of oocyte cryo-banking in order to allow proper patient counseling in the decision making
process in modern ART.
Materials and Methods: A search of published manuscripts and abstracts on human oocyte cryopreservation
was conducted in MEDLINE and other bibliographic databases. The genetics of oocytes, embryos, fetuses,
and newborns derived from cryo-thawed human mature (MII) oocytes tested by chromosomal studies werereviewed. In addition, perinatal data and follow-ups outcome of children conceived from such treatment wereanalyzed.
Results: The cryopreservation process may alter the oocyte spindle. However, in most survived oocytes thespindle apparatus has the ability to reform properly upon thawing and to recuperate its functionality. Oocytes
were investigated by preimplantation genetic diagnosis (PGD) of the 2nd polar bodies and by karyotyping
after fertilization and 90% were normal. The origin of the sperm used to fertilize the thawed oocytes wasreported not to affect the clinical outcome. The rate of embryonic aneuploidy, tested by PGD, is within the
expected range encountered in embryos derived from fresh oocytes. None of the aneuploidy found showed
any repetitive pattern. The number of live births derived from cryo-thawed oocytes has rapidly increased in
the last decade. Better post-thawing survival rates and encouraging pregnancy rates have been demonstrated
with vitrification as compared with slow freezing, and it became the method of choice worldwide. The rate ofcongenital anomalies is comparable to naturally and IVF/ICSI conceived children. Post-natal follow-ups
reported normal development.
Conclusions: No increase rates of aneuploidy or malformations were reported, and normal development was
found in post-natal follow-up in children born from cryo-thawed oocytes. While this data reassure the safetyof the clinical use of oocyte cryo-banking and is valuable in the decision making process in modern ART,
further prospective follow-up of the children is highly recommended.
◆Biosketch
Professor Ilan Tur-Kaspa is the President and Medical Director of the Institute for Human Reproduction (IHR),
Director of the Clinical IVF-PGD Program, Reproductive Genetics Institute, and Professor at the Dept of
OB-GYN at the University of Chicago. Prof. Tur-Kaspa is one of the world's most experienced Reproductive
Specialists involved in advanced IVF-ICSI treatment with PGD. He specializes in advanced ART,
reproductive imaging, and PGD. He is the author and co-author of over 90 scientific publications and bookchapters. He serves as a Reviewer for the leading Journals in Reproductive Medicine, such as Fertility and
Sterility, Human Reproduction Update and Reproductive BioMedicine Online. He is a co-Editor of the bookon Biotechnology of Human Reproduction. He was awarded the 2010 Star Award of the American Society for
Reproductive Medicine (ASRM) for over 10 years of continuous scientific contributions in the field ofReproductive Medicine.

143

●Concurrent Symposium C-9.3

What has happened on epigenetics in ART children?
Yukiko Katagiri
Toho University, Tokyo, Japan
In these years, an increase in imprinting disorders; such as Beckwith-Wiedemannsyndrome (BWS), Angelman syndrome (AS), and Silver Russell syndrome (SRS) in
children born after ART. Work on animal models suggests that in vitro culture may be a source of these
imprinting errors. Various clinical and experimental studies also suggested alterations of epigenetics of themale gamete, the female gamete, and embryogenesis in vitro culture to compare natural conception. Culture
conditions can also make a chance of imprinting alterations. ART has a possibility to change epigenetics withthe handling of gametes and early embryos as well as with in vitro culture because imprinted genes are likely
to be reprogramming at that times. It is also known that superovulation may effect on DNA methylation.
Oogenesis is one of an importance on epigenetics.
It needs to think here whether only ART gives the opportunity when epigenetics changes. There are thebackgrounds that increase risks of epigenetic alterations in couples requiring ART. Methylation of
spermatozoa in male infertile patients was different from in fertile men. For infertile couples with male factor,
in vitro fertilization or sperm injection can be used to assist conception; however, this could result inabnormally methylated sperm fertilizing an oocyte. In late years, with the change of the social background, thewomen needing infertility treatment has been ageing. We need to know that the expression of some genes wasaltered in ageing oocytes.
Though BWS and SRS are growth disorder diseases, some imprinted genes are associated with fetal growth.
In ART cases, imprinted gene expression and DNA methylation with human placental tissue were different
from neonates conceived naturally. It also has been known that epigenetic disorders are associated with some
adult diseases. The early life events in the development are very important for long-term health in their future.
ART was spread out very much in worldwide. However, there is not yet often reverse and others about thelong-term health of next generation. It is important epigenetic study and follow-up study in ART children.

◆Biosketch
Dr. Yukiko Katagiri obtained her specialty in Gynecology and Obstetrics and in Human Reproduction and
Genetics from Toho University. From 2001 to 2004, she had been in a research program in Cornell Institutefor Reproductive Medicine, Weil Medical College of Cornell University leaded by Dr. Gianpiero D Palermo.
She is Associate Professor in Department of Obstetrics and Gynecology, Toho University and Chief of the
Reproduction Center for women at Toho medical center, Omori Hospital in Tokyo, Japan. She is also Director
in Department of Clinical inheritance, Omori Hospital.

144

●Concurrent Symposium C-9.4

Pregnancy and Child Outcome after Assisted Reproductive
Technologies
Dov Feldberg
Helen Schneider Hospital for Women, Rabin Medical Center,
Jabotinsky St., Petah Tiqva, Israel
INTRODUCTION: One of the greatest achievements in the medical profession in the previous century was
the delivery of Louise Brown in 1978 and the first publication of Edwards and Steptoe in Lancet that year.
The second most important breakthrough in the field of Assisted Reproduction was the introduction byPalermo of Intracytoplasmic Sperm Injection (ICSI) procedure in order to solve the problem of severe Male
Factor Infertility. Until now about 4 millions of children were born from these novel technologies and the
major question that troubles the scientific world is Are ART Children healthy as their "brothers and sisters"that were born from natural conception?
MATERIALS AND METHODS: The prevalence of major congenital malformations including imprinted
gene defects was investigated in ART children compared with natural conception neonates that were treated
and born in our Hospital for Women between the years 1986-2006. We investigated as well all the reports on
major congenital malformations in ART children, compared to natural conception born children in the wholestate of Israel between the years 1997-2004. Our department is a part of Israel Birth Defects Monitoring
System and a part of the International Center for Birth Defects in Rome. We investigated the impact of the useof IMSI (Intracytoplasmic Morphologically Selected Sperm Injection) sperm selection technolopgy in severe
male factor infertility cases on the early abortion rate and prevalence of major congenital malformations in
ART neonates. All the global relevant literature concerning this crucial issue of congenital malformations and
ART was surveyed and extrapolated.
RESULTS: According to the results from our Hospital for Women, the Israeli Registry and the global
literature, the incidence of congenital malformations in ART children is nearly double as compared to naturalconception group. We could not present any specific malformations among the ART neonates and there wasno discrepancy in congenital malformation rate between standard IVF and ICSI micromanipulation children.
We found a possible linkage between ART children and increasing risk of Angelman andBeckwith-Wiedeman Syndromes that are associated with imprinted gene clusters. We present for the first timea significant improvement in neonatal outcome in terms of early abortion and congenital malformations rates
in IMSI children as compared to regular ICSI neonates.
CONCLUSIONS: There is no doubt that neonates of ART are at increased risk for major congenital
malformations including imprinted gene defects, as compared to natural conception neonates. No specificcongenital malformation was documented in ART children. No discrepancy was presented between standard
IVF and ICSI children. IMSI technology seems to reduce significantly the early abortion and congenital
malformation rates. This information should be presented to couples seeking ART treatment.
◆Biosketch
Born in Vilnius, Lithuania, 1947. Married with 2 children. Graduated Medical School of Hadassah andJerusalem University in 1971. Fellowship in Reproductive Medicine in 1984-1985 at the Yale New Haven
Hospital with Prof. Alan Decherney and Prof. Fred Naftolin. In 1985 establishes the fourth IVF Unit in Israel,at the Rabin Medical Center. Sabbatical in Reproductive Medicine in 1990-1991 at the Center for
Reproductive Medicine, Cornell University Hospital, New York, with Prof. Zev Rosenwaks. In 1993 receives
Professor degree at the division of OB/GYN Medical School at Tel-Aviv University. In 1994 appointed Vice
Chairman of OB/GYN Division, Rabin Medical Center. In 2001 appointed Acting Chairman OB/GYN
Division, Rabin Medical Center. From 2008 Executive Vice President of the World Association of145

●Concurrent Symposium C-9.4

Reproductive Medicine (WARM). About 120 publications in various journals mainly in the field ofReproductive Medicine. Several chapters in books and organization of National and International meetings.

146

●Concurrent Symposium C-10.1

Just Twis? Perinatal data from multiple pregnancies
Peter Brockerhoff
Department of Obstetrics and Gynecology,
Johannes Gutenberg-University Mainz, Germany
With the expansion of reproductive medicine worldwide the rates of multiple
pregnancies have considerably increased during the last decades. The rates of multiple pregnancies after
artificial reproduction differ enormously between the European countries. By data from literature as well as byan own analysis of the nations perinatal database it can clearly be demonstrated that twin pregnancies
compared to singleton pregnancies carry increased risks like pre-eclampsia, diabetes mellitus or perinatal
haemorrhage and increased fetal risks like prematurity, intrauterine growths retardation and higher perinatal
morbidity and mortality. In higher multiple pregnancies these risks occur with dramatically higher incidence
rates. Further collection of perinatal data from pregnancies after IVF or ICSI is on demand. Reproductive
methods therefore, which lead to higher rates of multiple pregnancies should be banned. Selective
Single-Embryo-Transfer ( eSET ) should be legalized also in those countries which still forbid this method by
their embryonic protection laws.
◆BiosketchPeter G. Brockerhoff , M.D.
29th of October 1948: born at Oldenburg, Germany
1955 – 1968 School education at Bremen
1968 – 1973 University of Heidelberg Faculty of Medicine and Faculty of Chemistry
Scholarship by the Students‘ Foundation of the German People
1971
Exchange student at Osaka/Nishinomiya , Japan ( sponsored by LIONS Club )1972
Exchange student at Tyler , Texas, USA
1973
Doctor‘s degree by the University of Heidelberg
1973
Marriage to Claudia Trommsdorff , High School Teacher
1974
Junior Doctor at Deptm. of Surgery University of Mannheim, Germany
1975 – 1984 Assistant Doctor of the Deptm. of. Obstetrics and Gynecology, Univ.-Mainz
1985 – 1993 Associate Professor and Consultant
Since 1994 Full Professor and Vice-Chairman Deptm. of Obstetrics and Genecology, Univ.of Mainz,
Germany
Board Member of the Medical Faculty of the University of Mainz
Board Member of the Senat Council for Education of the University of Mainz
Board Member of the Senat Board for Internationalisation University of Mainz
Member of the State Council for Medical Quality Control
Member of the Medical Association’s Ethic Board

147

●Concurrent Symposium C-10.2

Perinatal Outcomes of ART Pregnancies
Fethi Zhioua
IVF Center, Aziza Othmana hospital,
University of medicine, Tunis-TUNISIA
In-vitro fertilisation has been done for nearly 30 years; in developed countries at least
1-2% of births are from assisted reproductive therapy (ART). Perinatal outcomes, suchas preterm delivery, low birth weight and some obstetric complications seems to be increased after in vitro
fertilisation (IVF) compared with spontaneously conceived pregnancies (J.Hallyday.2004, Fujii.2010). But, it
is difficult to obtain accurate outcome information because of unmeasured confounders parameters. Besides,
some of the morbidity associated with ART does not result from the techniques but from the underlying healthrisks of being subfertile (Et.Reefhuis, 2009). So it is unknown whether IVF technologies or patient infertility
is the major contributor to adverse outcomes. Further, much of the amplified risk associated with ART isrelated to high birth order. Multiple births continue to be the major risk of couples needing fertility treatment.
Percent countries range from 25% to nearly 50%, with the rate of high order pregnancies as much as 40%.
The risk of birth defect after ART has been also discussed extensively in published studies. The subject
remains of concern, especially since the introduction of ICSI, with microinjection of single spermatozoa. The
overall prevalence of birth defects in the general population is approximately 4 %, with children born
following IVF/ICSI having a 25 – 40% higher risk of birth defects than children conceived spontaneously.
Further large population-based study are needed to explain specific risks.
There are many important deliberations for couples when considering the success of achieving a live birth andthe adverse outcomes potentially associated with ART. The most accurate and up-to-date information possible
should be given to them, along with counselling to help with understanding of the potential adverse outcomes.
Only then can couples make the choice about IVF that may be best for them.
◆Biosketch
PRESENT ADDRESS:IVF Center. Hopsital of Aziza Othmana, Place de la kasbah Tunis, Tunisia
EDUCATION AND TRAINING:
Presidency of the department of gynecology and obstetrics (2004 - 2005)
Professorate degree in gynecology obstetrics: University of medicine Tunis 1992
Medical doctorate in gynecology obstetrics: University of medicine Tunis 1986
PROFESSONAL ACTIVITIES:
Several national positions including:Presidency of the National society of gynecology and obstetrics (2008-2011)
Member of the National Committee of reproduction (2005 - 2011)
Counselor of the Minister in gynecology and obstetrics (2004-2011)
Member of ESHRE since 1992
Member of ASRM since 1998
AREAS OF INTEREST:
Gynecology and Obstetrics include infertility, high-risk pregnancy and gynecologist surgery
Biomedical ethics
Aneuploidy screening and PGD

148

●Concurrent Symposium C-10.3

Outcome of IVM babies
Ri-Cheng Chian
Associate Professor, Department of Obstetrics and Gynecology
McGill University, Montreal, Canada H3A 1A1
Introduction: Initial oocyte in vitro maturation (IVM) treatment was performed for women infertility withpolycystic ovary syndrome (PCOS), and then IVM technology was expanded to treat the over responders andthe poor responders for gonadotropin stimulation. It has been estimated that more than 2,000 babies born with
IVM treatment in the worldwide, and the clinical pregnancy and implantation rates were approximately
30-35% and 10-15% per embryo transfer, respectively (Chian et al., 2004). Although it has been believed that
IVM treatment is not associated with any additional risk in terms of obstetric outcomes and congenitalabnormalities (Buckett et al., 2007), it was based on a small number of live births at one fertility center. In thisstudy, we performed a large scale survey for IVM babies born in the worldwide.
Methods: Data were collected from the worldwide with where IVM babies born. Congenital abnormality,
gestational age, birth weight, Apgar scores, cord pH, growth restriction, pregnancy complications, mode ofdelivery, and multiple pregnancies were analyzed.
Results: A total of 1,421 IVM babies born world widely were analyzed. The data showed that congenitalabnormality, Apgar scores, cord pH, growth restriction, and pregnancy complications were comparable withthe control database.
Conclusions: Based on the data collected, it seems that IVM treatment is not associated with any additionalrisk in terms of congenital abnormalities for the IVM babies.
◆Biosketch
Currently, Dr. Chian is Professor (rank as Associate Professor with Tenure) at Division of Reproductive
Biology, Department of Obstetrics and Gynecology, McGill University, Montreal, Canada.
Dr. Chian is the key person developed in vitro maturation (IVM) of human oocytes for clinical application andresponsible for cryopreservation of human oocytes produced pregnancies and live births at McGill University
Health Center (MUHC) in Canada. Dr. Chian published numerous research papers and presentations inrefereed journals and international/regional conference. Dr. Chian edited two books, and contributed many
book chapters. As a guest or invited speaker, Dr. Chian delivered numerous lectures in different societies andcountries.
Dr. Chian was former Associate Editor for the journal, Human Reproduction and is an Editorial Board
Member for Journal of Assisted Reproduction and Genetics and other two journals. Dr. Chian is acting as areviewer for many other scientific journals, including Fertility and Sterility, Reproduction, Biology of
Reproduction, Reproductive BioMedicine Online, Endocrinology, The Journal of Clinical Endocrinology &Metabolism, and The Lancet, etc.
Dr. Chian‘s research interests are focused on 1) Ovarian function; 2) The mechanism of oocyte maturation andactivation; 3) Fertility cryopreservation.
Dr. Chian speaks English, Chinese, Korean and Japanese as well as some knowledge of French.

149

●Concurrent Symposium C-10.4

Perinatal outcome of vitrified human blastocysts in 11 years experience(5434 attempted cycles) including the rate of monozygotic twinning(MZT)
Tetsunori Mukaida1, Chikahiro Oka2, Tetsuya Goto2,
Katsuhiko Takahashi1
1
Hiroshima HART Clinic, 2Tokyo HART Clinic
Introduction: Vitrification has been recognized as a useful method for cryopreservation of human blastocysts(BLs). However, a possible drawback of vitrification could be the use of a relatively high concentration of
cryoprotectants, which may affect the embryo and subsequent development. Also, increased incidences of
MZT related to BL transfer including vitrified BL compared with natural conceptions have been reported.
Therefore, we analyzed reproductive outcomes and perinatal incidences after 13 weeks of gestation in infants
born after vitrified BL transfer.
Materials and Methods: Clinical and perinatal outcome of vitrified BL transfer was summarized between
Jan. 2000 and Dec. 2010. Mean age was 36.4 years. Procedures of vitrification were carried out at 37 0C
containing 7.5% DMSO and 7.5% ethylene glycol (EG) and 15% DMSO, 15% EG, 1% Ficoll 70 and 0.65 M
sucrose. For cooling, the BLs were loaded on a small nylon loop and plunged directly into LN2. Warming was
performed by placing the tip of the cryoloop into 0.5 M sucrose for 2 min and then 0.25 M sucrose for 3 min.
For expanded BLs, the blastocoelic cavity was artificially collapsed prior to vitrification, and zona hatching
was performed after warming with laser pulses Survival was assessed based on the morphological integrity
and re-expansion of the blastocoele. Embryo transfer was performed 2-3 hours after warming. The uterine
endometrium was prepared with exogenous E2 and P4. Pregnancy was defined as the presence of gestationalsac(s) (GS), and MZT was defined as two fetus with fetal heart beat in the same GS.
Results: A total of 8960 vitrified BLs from 5434 cycles were warmed and 8432 survived (94.1%). Transfer
was cancelled in 87 cycles (1.8%), since warmed BLs did not survive or were not suitable for transfer. In 5437
transfers, 2568 cycles resulted in pregnancy (48.0%), and 686 cycles (26.7%) ended with miscarriage. Seven
thousand, five hundred and thirty two vitrified BLs were transferred and 2882 were implanted (38.3%). Mean
number of BLs transferred per cycle was 1.41. Two hundred two (13.6%) were multiple pregnancies.
Seventeen hundred and seventy one children were born in 1553 deliveries. Forty one clinical pregnancies
were confirmed as MZT (2.6%). Cesarean section was performed in 804 deliveries, and mean gestational age
was 38.4 weeks. No bias in the sex ratio was observed with 909 boys and 862 girls. Mean birth weight was
2819g. Thirty two babies in 32 deliveries had either congenital birth defects or perinatal complication (1.9%),
including six chromosomal abnormalities (one 18 trisomy, three 21 trisomy), three multiple anomaly, one
death due to hydropus, four stillbirth of unknown cause (25, 29, 30, 37 weeks of gestation), one anencephaly,
one spina bifida, nine congenital heart malformation and three minor anomaly in hands and/or foot, onecongenital esophageal, and one biliary duct obstruction, one Cornelia De Lange syndrome (CdLS), and one
Treacher Collins syndrome.
Conclusion: Clinical results showing above section as well as the incidence rate of MZT (2.6%) and rate ofcongenital defect and neonatal complication (1.9%) were both similar to fresh BL transfer as we reported
previously (2.3% & 2.0%) confirm that vitrification of BLs using the cryoloop technique are effective,
practical, and confirming the safety of our procedure. Also, the high implantation rate encourages us toestablish single BL transfer.
◆Biosketch

150

●Concurrent Symposium C-10.4

He completed his board certified OB&GYN training in OB&GYN dept. at Kochi Medical School and
Hospital, Japan in 1990. During that training, he studied abroad to the Univ. of Miami, School of Medicine, inthe Biochemistry and Molecular Biology Dept. After the training in Japan, he became a lab director in a
private fertility clinic, the Diamond Institute for Infertility in New Jersey, USA until 1995. For the last 15
years, he became co-director of the Hiroshima HART Clinic. His main interest is vitrification for gametes andembryos: he is one of the pioneers for vitrification including reports of successful birth of day 2-3 vitrified
embryos with straw (1998), and the vitrified blastocysts using a cryoloop (2000). His original video for
vitrification was received the prize in Video: ART Category in 61st ASRM. His abstract of perinatal outcome
in vitrified BL program in 11 years experiences was pre-selected for the Clinical Science Award for oralpresentation at the 2010 ESHRE in Rome.

151

●Concurrent Symposium C-11.1

Novel embryo culturing and monitoring systems
Artur Bernard1, Peter Kovacs1, Szabolcs Matyas1,
Csaba Pribenszky2, Gabor Vajta3, Steven Kaali1
1
Kaali Institute, IVF Center, Budapest Hungary,2
Dep.of Animal Breeding and Genetics, Faculty of Veterinary Science,
Szent Istvan University Budapest Hungary,
3
BGI Ark Biotechnology Co. Ltd Shenzhen, Shenzhen, China
In vitro culture of mammalian embryos may be remarkably successful when the embryos are kept in relatively
large groups during the whole culture period. Although preimplantation stage embryos are autonomous
organisms, their developmental competence is considerably increased by factors secreted by other embryos
cultured in close vicinity. The selection of the best embryo(s) for transfer is of utmost importance, to achieve
high pregnancy rates and to avoid multiple pregnancies. Most selection methods require individual culture ofembryos. Very recently, systems have been developed to meet both, seemingly conflicting requirements. The
group or communal effect can be mimicked by culturing embryos in small amount of media. In these systems
paracrine effects may be replaced by autocrine ones, ligands produced by a single embryo may stimulate theembryo proper. Another possible benefit of decreasing the amount of culture media is that the embryo may
establish and maintain a microenvironment, a mixture of physical, chemical and biological factors that support
appropriately metabolism and development. A similar situation may occur in the virtual space of the oviduct;
however, the high medium / embryo rate of most in vitro culture systems may require disproportional effort
from embryos to accommodate continuously to the suboptimal conditions that are artificially provided.
Microwells produced on the bottom of the wells of the culture dishes (well of the well, WOW system) may
provide an appropriate solution. The quantity of medium in the well is minimal, allowing accumulation of
autcrine ligands and establishment of the appropriate microenvironment. On the other hand, the free contact
with a large amount of medium outside the microwell allows exchange of components by slow diffusion,
hampers accumulation of potentially dangerous end products of metabolism, and ensures supply of nutrients.
Application of such micro wells has considerably improved in vitro development of both zona-intact andzona-free bovine and porcine embryos, human zona intact, in vitro produced embryos, and increased
pregnancy rates after transfer. Versions of the WOW system can also be applied for monitoring embryo
metabolism as well as morphological features of individual embryos during the whole period of development.
With proper arrangement of wells, a single objective can monitor 9 or more embryos in a single field of view.
Recently, a compact time-lapse microscope / camera system (Primo Vision; Cryo-Innovation, Budapest,
Hungary) has been developed to exploit this possibility, while avoiding the usual complicated structure andpotential harmful effects of time-lapse instruments. Experiments performed in mouse have proved the value ofthe system, the time of the first and second cleavage strongly indicates the probability of blastocyst
development, while occurrence of fragmentation - even if completely reabsorbed in hours - markedly decrease
the developmental competence. This system has resulted in the first baby in the world born after time-lapse
investigation, followed with hundreds worldwide. The combination of WOW and compact time-lapse system
offers new perspectives for improvement and proper monitoring of cultured embryos.
◆Biosketch
Artur Bernard obtained his MD degree (1969) specializing in Obstetrics and Gynecology (1974) and earned a
PhD degree in human reproduction (1991) in Hungary. In 1978-79 he spent more than one year as a Lalor
Fellow at Washington University, St. Louis, Mo. USA. Between 1987 and 1990 he worked in Germany, one
year in the IVF Team of the University Erlangen, and later in Dusseldorf. 1993 he has been appointed as
Visiting Associate Professor in the Dep. of Ob/Gyn. at the Albert Einstein College of Medicine, Bronx, New
York. Since 1992 he has served as the scientific director of the Kaali Institute IVF Center in Hungary.

152

●Concurrent Symposium C-11.2

Effects of cumulus cells removal after 6 h co-incubation of gametes on
the outcomes of human IVF
Guoning Huang
Chongqing Reproductive and Genetics Institute
OBJECTIVE: To investigate the effects of cumulus cells removal after 6 h
co-incubation of gametes on the fertilization, polyspermy, multinucleation and clinicalpregnancy rates in human IVF.
MATERIAL AND METHODS: A total of 1200 IVF-ET cycles undergoing 6 h co-incubation of gametes in2009 were included in this study. Inclusion criteria were: female age younger than 38 years, first IVF
treatment, with bi-ovary and normal ovarian response, e.g., 4~20 oocytes can be obtained. A 6 h period of
co-incubation was applied in all IVF cycles. According to the history of infertility, cumulus cells weremechanically removed either 6 h post-insemination or 20 h post-insemination. For couples with primaryinfertility, or unexplained infertility, or mild oligospermia or asthenospermia, the cumulus cells were removed
at 6 h of insemination for the polar body observation (6 h group, n=565). Of which, 80 cycles received early
rescue ICSI due to fertilization failure or low fertilization rate at 6 h of insemination. For couples withsecondary infertility and normal semen analysis, the cumulus cells were removed at 20 h of insemination as
routine (20 h group, n=635). Of which, 3 cycles received late rescue ICSI due to fertilization failure at 20 h ofinsemination. Fertilization, polyspermy (more than 3PN), multinucleation and clinical pregnancy rates werecompared between the two groups (rescue ICSI cycles were not included in the comparison in both groups).
RESULTS: Significant difference (Pless than 0.05) was observed between the two groups regarding
polyspermy rates (7.48% in 6 h group and 9.22% in 20 h group). No difference was observed between the twogroups regarding normal fertilization rates (2PN rate) (64.89% in 6 h group and 65.74% in 20 h group). No
difference was observed between the two groups regarding multinucleation and clinical pregnancy rates(11.01% and 65.15% in 6 h group, 10.75% and 66.93% in 20 h group, respectively). The clinical pregnancy
rate was 51.43% in cycles receiving early rescue ICSI, while no clinical pregnancy was obtained in cycles
receiving late rescue ICSI.
CONCLUSION: The present results indicate that cumulus cells removal at 6 h of insemination is a relatively
safe operation, which yielded comparable normal fertilization rate, multinucleation and clinical pregnancy
rates compared with 20 h group. This protocol may be beneficial for early obsevation of fertilization failure
and make early rescue ICSI possible.
◆Biosketch
Guoning Huang is the professor and director of Chongqing Reproductive and Genetic Institute in Chongqing
Obstetrics and Gynecology Hospital. He is the Secretary-General of Chinese Society of Reproductive
Medicine.

153

●Concurrent Symposium C-11.3

An integral view on therapeutic options and lab-techniques forindividualized therapy in IVF
Nicolas Zech, M.D., PhD
IVF Center Prof. Zech - Bregenz
In the early days of IVF the consultation of patients was a rather easy task with only
limited options available. Nowadays, there are various – sometimes very complex - treatment modalities
available to improve the individualized IVF outcome of which all should be discussed with the couple.
This presentation will give an overview on new treatment modalities and lab-techniques which continuously
evolve with sometimes radical changes in the approach on how patients are being consulted and treated aswell as the challenges and complexities we face as a team to guarantee the highest individualized chance for asafe conception.
◆Biosketch
Univ. Doz. Dr. med. Nicolas Zech studied medicine at the Medical University Innsbruck, Austria. His
research interests have focused on stem cells since completing his thesis on expansion of mobilized peripheral
blood progenitor cells. At the IVF Centers Prof. Zech he focused on in-vitro culture, embryology andmicromanipulation techniques. He has been trained as Obstetrician and Gynecologist at the University
Hospital Zurich, Switzerland, and has finished his specialty in OB/GYN in 2005. Nicolas holds a Diploma for
Genetics from the Austria Society for Medical Doctors. He was for 2 years on a research fellowship at the
Reproductive Genetics Institute in Chicago (Yury Verlinsky – Pioneer of Preimplantation Genetic Diagnosis),
USA. Habilitation (Venia docendi) in Obstetrics, Gynaecology, Reproductive Medicine and Endocrinology at
the Medical University Graz 01/2010 - Topic: Stem Cells and Assisted Reproductive Technologies - Title:
Characterization, isolation/ establishment, culture and storage of clinical grade stem cells with input fromassisted reproductive technologies. Nicolas joined the Chief Executive Program at Harvard Business School(HBS) in August 2008. He is a Harvard Business School Alumnus (Owner-President-Management Program OPM41) since March 2011. He was winner of HBS-OPM41 Idol in 2009. He is Medical Director at the IVF
Center Prof. Zech - Bregenz.

154

●Concurrent Symposium C-11.4

How to manage the modern IVF laboratory
Zsolt Peter Nagy
Reproductive Biology Associates
Introduction. Managing appropriately a contemporary IVF laboratory is one of themost complex and challenging work, that is critically important to achieve success in
assisted reproduction treatment. The objective of the present submission is to provide
a review of the most important contributing factors to manage an IVF Laboratory successfully. Laboratory
design is the cornerstone of a well functioning laboratory, usually it can be considered when setting up a newlaboratory or when moving to a new space. It is critical to assign a sufficiently large enough space (relative tothe expected work volume) that provides conditions to a seamless flow of procedures. High quality ofmaterials with easy to clean surfaces are required to build out the space. HVAC system providing highpressure of clean air of all time is a principal condition. Automated gas systems (providing N2, CO2 and gas
mix/triple gas) with highest purity gases are essential, with alarms and sufficient back up. There are plenty ofchoices of different instruments, most importantly, microscopes, micro-manipulators and incubators,
important that those chosen are high quality and reliable. Incubators should be able to provide conditions to
low O2 culture conditions. Disposables that are used in the IVF lab should be specifically manufactured tofulfill related requirements and be approved for routine use. Another most critical component of the laboratory
work is the culture system, the choice of culture medium, protein source and oil. The embryo culture systemshould be established in a way to provide conditions for successful extended culture as well. Those items usedin the IVF lab should be obtained from trusted sources but independently, they should be tested before regular
use to verify appropriateness. Quality control and quality assurance systems must be in place and should be
used rigorously to verify for appropriateness all components of the laboratory, including daily checks on all
critical components of the lab. Laboratory personal has to be appropriately trained having both adequate
theoretical knowledge and procedural skills. Regular evaluation on individual performance is critical, as wellas continuing update on knowledge and skills. Team work within the laboratory and with clinicians as wellwith other supportive staff is essential. Participation in proficiency testing and laboratory inspection by
independent third party organizations is highly recommended.
Conclusions. There are several hundreds of variables that are constantly influencing laboratory performance,thus to manage it successfully it requires a well established system and well trained, knowledgeable
professionals.
◆Biosketch
Dr. Nagy obtained his MD (1986) and his Ob&Gyn degrees (1996) at the Semmelweis Medical University in
Budapest. His PhD was granted at the Free University of Brussels in 1997, where he worked in the team thatdeveloped the ICSI procedure. More recently, he has contributed to the development of a highly efficient
oocyte cryopreservation procedure that gave bases to establishing the first donor cryo-bank in USA. He is
author of more than 150 publications and book chapters. Dr. Nagy serves on the board of several different
societies; as well he is Associate Editor for Human Reproduction and RBMonline. Currently, Dr. Nagy is the
Scientific and Laboratory Director at Reproductive Biology Associates, Atlanta, USA.

155

●Concurrent Symposium C-12.1

Chemical substances exposure on IVF-ET environment
Tsunehisa Makino
Tohbu Hospital, Gotemba City, Shizuoka, Japan
Introduction: Very little has been discussed on chemical substances exposure on
ART environment, although recent estimates have indicated that more than
50,000,000 chemicals exist in the global environment.
Materials and Methods: Several candidate chemicals were selected from lists of chemicals previously
reported to have the potential or capability of inducing possible health risks and/or congenital disorders.
Sensitive and specific quantitation methods have been developed for the measurements of these chemicals,
which included phthalates (DEHP,MEHP) and perfluorinated compounds (PFCs), polybrominated diphenyl
ethers (PBDEs), nonylphenol (NP), bisphenol A (BPA), nicotine and its metabolites, organophosphate andpyrethroidal insecticides, and heavy metals. Peripheral blood and urine samples from mothers, cord blood,amniotic fluid, and breast milk samples were collected without manual contamination and the levels of theselected chemicals in these various samples were measured. After establishing reference standards forchemical contamination on human specimens, especially during the perinatal period, we investigated how
these endogenous amounts of chemical substances might induce health hazards in the next generation by
analyzing epigenetic alterations in mouse embryonic stem cells (ESCs) and in part in human induced
pluripotent stem cells (iPS). We also analyzed the levels of DEHP,MEHP,PBDEs, Np, BPA,PFCs in culture
media used for assisted reproductive technology (ART).
Results and Discussion: Though the genetic bases of physiological variations as well as those of many
pathologic abnormalities are rapidly being elucidated, hereditary diseases and congenital malformations arestill major concern in mordern obstetrics. In the present study, the concentrations of chemical substances in
the fetomaternal eivironment, as detected using newly developed and very sensitive and specificchromatographic and spectrometric asay systems, were relatively low (approxomately 1 - 10 ppb) and were
far from the pharmacologic doses utilized in teratogenic or toxic studies in animals. However wedemonstrated that 0.1ppb of DEP (diethyl phosphate, a metabolite of organophosphate insecticides), cotinine(a metabolite of nicotine), mercury (Hg), selenium (Se) or S-421 (octachlorodipropyl ether, a synergist of
insecticide) and 1 ppb level of MEHP and deca BDE (brominated diphenyl ether) are capable of causing
significant epigenetic changes. The selection of an appropriate medium for each step in gamete preparationand zygote cleavage is widely known to have a crucial impact on the success of implantation during in vitro
fertilization. We investigated varous kinds of IVF media, including sperm and oocyte buffers and media,
culture oil, fertilization and cleavage media and blastocyst media, and detected 10 - 100 times higher levels of
MEHP and PBDE in some groups, indicating that some IVF media are highly contaminated by thesesubstances.
Conclusion: Several environmental chemical substances at doses detectable in fetomaternal specimens andIVF media can induce significant alterations in the epigenetic profile. The present study also showed that our
method using mouse ESCs and human iPS cells to assess epimutagens is simple and sufficiently sensitive toevaluate the toxic and/or teratogic influences on environmental chemical substances. (This study was
supported by a Health Science Reserch Grant from the Ministry of Health, Labour and Welfare, Japan)

156

●Concurrent Symposium C-12.1◆Biosketch
Dr. Tsunehisa Makino was graduated from School of Medicine, Keio University, Japan, 1964. Dr. Makino has
been a postdoctoral fellow at The Laboratory of Human Reproduction and Reproductive Biology, Harvard
Medical School between 1970-1973 and was Assistant Professor of the Department of Obstetrics andGynecology, Harvard Medical School in 1973. Dr Makino promoted his position to Professor and Chairman,
Department of Obstetrics and Gynecology, Tokai University Hospital, Japan, in 1995. Dr. Makino waschairman of the Executive Board Meeting of the Japan Society for Immunology of Reproduction (JSIR), also
the president of IVth Conference of the Pacific Rim Society of Fertility and Sterility, held in Okinawa, andIXth International Congress of Reproductive Immunology held at Hakone, Japan, in 2004. Dr. Makino has
been the director of Tohbu Hospital, At Gotemba City, Shizuoka, Japan, since 2005.

157

●Concurrent Symposium C-12.2

Recent insights into the oocyte maturation in ART
Tae Ki Yoon, M.D.
Fertility Center of CHA Gangnam Medical Center,College of Medicine, CHA University, Seoul, Korea
Gonadotropin stimulation for human in vitro fertilization-embryo transfer (IVF-ET)
program has been used to retrieve multiple numbers of mature oocytes, subsequently leading to increased
number of embryos to be transferred and the pregnancy rate. However, this regime has potential disadvantages
such as ovarian hyperstimulation syndrome, especially in women suffering from polycystic ovarian syndrome(PCOS). In vitro maturation (IVM) followed by IVF (IVM-IVF program) could be an alternative to bypass
disadvantages of conventional IVF and give better chances to achieve pregnancy to patients suffering frominfertility. Recent works have clearly showed that unfavorable conditions that could occur during growth andmaturation of immature oocytes greatly affect embryo development and further clinical outcome. Thus, age,
etiology, menstrual cyclicity of patients could influence maturity and growth as well as the number of
retrieved oocytes in ART program. Furthermore, follicle size, oocyte diameter, and hormonal environments
have an impact on cytoplasmic maturation of oocytes. With respect to advance of IVM-IVF programs, it isnecessary to investigate underlying mechanisms and regulatory factors which contribute to achieving higher
efficacy of IVM as well as in vitro culture system. Composition of culture media and other laboratory factors
such as hormones, growth factors, energy substrates, and scavenger system have been continuously fine-tuned
to improve nuclear and cytoplasmic maturation of immature oocytes in vitro. Increased number of oocyteswith good quality for IVF-ET by employing an optimized IVM system would provide practical benefits topatients with PCOS, POF and repeated IVF-failure. Moreover, it could reduce an ethical burden about thedisposal of retrieved oocytes with low developmental competence. Current knowledge and achievements in
IVM of human oocytes for clinical application should be further expanded to enhance clinical outcome in
IVM-IVF-ET programs.
◆Biosketch
NAME:
OFFICE ADDRESS:

Tae Ki Yoon, M.D.
Department of Obstetrics and Gynecology, CHA University
Fertility Center of CHA Gangnam Medical Center,
650-9, Yeoksam-1-Dong, Gangnam-Ku,
Seoul, 135-081, Korea

EDUCATION:
1969 - 1971 Premedical College, Yonsei University, Seoul, Korea
1971 - 1975 Medical School, Yonsei University, Seoul, Korea
1983 - 1986 Department of Medical Science, The Graduate School, Yonsei University, Seoul, Korea
POSTGRADUATE TRAINING:
1978 - 1979 Intern, Paik Hospital, Inje University, Seoul, Korea
1979 - 1983 Resident, Department of Obstetrics and Gynecology, Severance
Hospital, Yonsei University, Seoul, Korea
1988 - 1989 Postdoctoral Fellow, Reproductive Endocrinology and Infertility
Department of Obstetrics and Gynecology, Yale University, New Haven C.T., USA

158

●Concurrent Symposium C-12.2

PROFESSIONAL POSITIONS HELD:
2009 - 2011 President, Korean Society of Assisted Reproductive Technologies (KOSAR)
2009 Member of Korea National Bioethics Deliberation Committee
2004 - 2008 Chair of Committee Member, Technical Expert of Artificial Fertilization,
Korean Bioethics Review Board
2004 - 2008 Chairman of ART Subcommittee, Korean Society of Obstetrics & Gynecology
2004 - 2008 Medical Treatment Consultant, Korean Broadcasting System
1997 - 2008 Professor and Chair, Department of Obstetrics and Gynecology
School of Medicine, CHA University
2011 Professor and Dean, School of Medicine, CHA University
1990 Director, Fertility Center, CHA Gangnam Medical Center
1983 - 1987 Clinical Staff, Department of Obstetrics and Gynecology
AWARDS, HONORS & DISTINCTIONS: 11 times (International and Domestic)
PUBLISHED BOOKS AND CHAPTERS: 10 times (International and Domestic)
MEDICAL JOURNALS: 52 publications (International, SCI journals), 120 publications (Domestic journals)
INVITED FACULTY AT INTERNATIONAL CONFERENCES: More than 25 times

159

●Concurrent Symposium C-12.3

Use of high concentrations of cryoprotectants: is it a justified argument
to prefer slow freezing instead of vitrification?
P. Vanderzwalmen1,2,4, B. Wirtleitner1, B. Lejeune2,
S. Vanderzwalmen2, F. Ectors4 , N. Zech1
1
IVF Centers Prof. Zech - Bregenz, Austria
2
Centre Hospitalier Inter Régional Cavell (CHIREC), Braine l‘Alleud,
Belgium
3
GIGA-Research, University of Liège, Liège, Belgium
Introduction
The use of high levels of cryoprotectants (CPs) in solutions applied to vitrify oocytes or embryos is anargument to still prefer slow freezing procedure. Is it a justified argument?
Material and Methods
Cinematographic analysis of the change in volume in relation with different concentration of cryoprotectants
Conclusions
Out of three studies using mice zygotes we may assume:- The intracellular concentration of CPs is far lower than the one in the vitrification solutions- The intracellular concentration of CPs in the vitrified zygote is in contrary to the common beliefs even lower
than the one observed after a slow freezing procedure- Survival after slow freezing reflects the presence of an intracellular vitrified state in these cells◆Biosketch
Pierre Vanderzwalmen, embryologist, is currently working as scientific coordinator at the IVF center of Prof
Zech in Austria and in the CHIREC IVF institute of Prof Lejeune in Brussel.
His scientific investigations focused: on the relationship between selection of spermatozoa and embryo
selection criteria on the outcome of embryo development, and on vitrification of oocytes and embryos.

160

●Concurrent Symposium C-12.4

Embryo culture: Can we perform better than Nature?
Gábor Vajta1, Aniko Reichart2, Laura Rienzi3
1
BGI Shenzhen, Shenzhen, China,
2Forgacs Institute, Budapest, Hungary,
3
G.EN.E.R.A Centre for Reproductive Medicine, Rome, Italy
Culture of preimplantation stage embryos has always been a key element oflaboratory embryology, and contributed substantially to the success of many assisted reproductive procedures.
During the past decade, its importance has increased even more, as extended in vitro embryo culture andsingle blastocyst transfer have become indispensable parts of the approach to decreasing the chances ofmultiple pregnancies while preserving the overall efficiency of the treatment. Based on the impressive
achievements during the past decade there is an increasing, although rarely declared doubt if the present
efficiency of in vitro embryo culture can be considerably expanded in the future. The basis of this opinion isthat there are natural limits of advancement, and essentially we cannot cross the laws of Nature. However, inspite of the scientific and commercial challenge stimulating research worldwide to optimize embryo culture
conditions, a consensus is missing even in the basic principles including composition and exchange of media,
the required physical and biological environment, or even the temperature of incubation. Basic elements ofembryo culture are derived from somatic cell cultures and have been applied mechanically in embryology,
disregarding the special needs of preimplantation embryos. Unfortunately most producers of commerciallyavailable media fail to provide the exact composition of their products, hampering further development, as any
comparison can only be made between different brand names instead of constituents. In this presentation wesummarize the controversies, demonstrate the fragility of some widely accepted dogmas, and try to generate
an open-minded debate towards rapid and efficient optimization. New approaches expanding the traditionalframes of mammalian embryo culture are also discussed including application of new culture devices(microwells, or microchannel-microfluidic system), the need of selection of the right animal model for testing
devices and media, and the optimal balance between undisturbed quiet environment and well-defined stresstreatment to improve developmental competence. Authors are confident that substantial improvement may beachieved that may expand considerably the possibilities of future assisted reproduction in humans.
◆Biosketch
Gábor Vajta has obtained M.D. degree (1976) specialty degree (1979) and Ph.D. (1988) in human pathology
in Budapest, Hungary. In 1990 he has started to work in mammalian embryology. In 1999, he obtained a DSc
degree in Copenhagen, and is now Affiliated Professor of the University of Copenhagen, Honorary Professorof Beijing Institute of Genomics, and Adjunct Professor of James Cook University, Australia. He was authorof more than 160 scientific publications and innovator in 5 patents His incubation, embryo culture,
cryopreservation and nuclear transfer methods are known and used worldwide. He has led the research groupthat produced the first cloned animal of Africa as well as that of Scandinavia, and in collaboration with his
Chinese and Danish colleagues he contributed in establishment of the first porcine model in the world for the
Alzheimer disease. His current work includes propagating/teaching innovative new technologies in the field ofdomestic animal and human embryology.

161

●Concurrent Symposium C-13.1

Superovulation/IUI - An Alternative to IVF
Bill Yee, M.D., F.A.C.O.G.
Long Beach, CA
Infertility affects 12-15% of couples the United States, but only a very small
percentage will seek IVF because of its cost. The cost of an IVF cycle, including
medications, range from $12,000 - 15,000 USD. Of the couples undergoing IVF,
over 50% of the women have patent tubes and over 75% of the men have motile sperm totaling more than 10
million. Superovulation/IUI, where gonadotropin is used to increase the number of eggs, is not only a viableoption but also an attractive financial alternative instead of IVF for these couples. The cost of an IVF cycle isfive to six times more than a SO/IUI cycle, which is typically only $2,000 - 3,000 USD. Compared to IVF, thesuccess rate with SO/IUI is only about one-third less.
A typical SO/IUI cycle consists of using 75 to 150 IU of gonadotropin for 8 to 10 days. When two follicles
reach a mean diameter of 18-20 mm, hCG is usually given and a single insemination is performed 36 hours
afterward. In a series of over 5,500 SO/IUI cycles performed this way, in women less than 35 years old andwith more than 10 million motile sperm, the clinical pregnancy rate was 21%.
The success rate was higher in a subset of patients who had two inseminations instead of one and when hCG
was given at a much larger follicular size. The pregnancy rate in this group was 29%, compared to 53% in thesame age group of women undergoing IVF.
The methodology and the rational for this approach, along with a probable reason that would account for suchhigh success rate with technique will be discussed.
◆Biosketch
Bill Yee, M.D., the Medical Director of Reproductive Partners Medical Group, Inc., started the In Vitro
Fertilization (IVF) program at Long Beach Memorial Medical Center in 1986. Dr. Yee is internationallyrecognized for his expertise in the field of Reproductive Medicine.
His work resulted in the first successful frozen embryo transfer resulting in a healthy baby in the United States.
Dr. Yee is well known for his expertise in microsurgical, laparoscopic, and hysteroscopic surgical techniques.
He is the lead author of Transvaginal Sonography in Infertility and co-authored Atlas of Gynecological
Surgery. His pioneering IVF work also produced the first frozen embryos in the lowland gorilla.
Recently, he completed a three year term on the Board of Directors of The American Society for Reproductive
Medicine (ASRM), the national and international leader in multidisciplinary information, education, advocacy
and standards in the field of reproductive medicine.
Dr. Yee also served on the RESOLVE National Board of Directors. RESOLVE is the National Infertility
Association, established in 1974, dedicated to providing education, advocacy and support to men and womenfacing infertility issues.
Dr. Yee‘s commitment to quality IVF care has led to the positions of President of the Society for
Reproductive Technology (SART), Deputy Commissioner of the College of American Pathologists (CAP) for
Reproductive Laboratory Accreditation, and Director of the American Society for Reproductive Medicine.
Numerous publications have recommended him as one of this country‘s top doctors, including Ladies‘ Home
Journal, Good Housekeeping, and Top and Best Doctors in America.
He is the longest sitting member on the Executive Council of SART, since 1994. Dr. Yee also confounded thePacific Rim Society for Fertility and Sterility.

162

●Concurrent Symposium C-13.2

Nurturing the ovaries: Treatment of patients with high FSH withdelayed stimulation using low dose gonadotrophins
MKH Leong, AKN Doo
The Womens Clinic, 318 Central Building, 1-3 Pedder Street,Hong Kong and the IVF Centre, Hong Kong Sanatorium and Hospital,
Hong Kong
One of the major problems in IVF has been that of patients who are extreme
poor responders. These patients are identified by their previous history, by raised
FSH, low antral follicle count, low AMH, or a combination of some or all of these. Probably the commonest
criteria of diagnosis are low AFC and raised FSH.
FSH levels over 12 or 15 U/L is generally the cut off level as raised. Over these levels, these patients are
excluded in any 'normal' IVF studies. They are considered 'abnormal' and this group of patients also
commonly face canceling their cycle, and denied an IVF treatment. Most of these patients are older also, andso not to treat them in consideration of IVF outcome amounts to a humiliating decision. In present times,
especially in the Western world, these unfortunate couples are usually offered egg donation (sometimes
forced) and dissuade from routine IVF. Egg donation has its problems, genetic, social, and psychological, andmay not be acceptable to a lot of couples. A better solution has to be found.
We looked at this particular problem, and asked "why not?" - these are cycling patients, most of them quite
regularly. Our reasoning is this: if they are cycling, they must produce estrogen sometime during their cycle,
and this would be from the granulosa cells lining the follicles. In other words, follicles must develop to make aregular cycle. They are not responsive to especially high doses of FSH in the beginning because their follicles
are not ready to respond, the receptors are not ready; and they are producing high doses of FSH by their own
Pituitary gland already. But, given the right timing, we should be able to take over from when their own
secretion of FSH drops off by adding exogenous FSH to continue follicular maturation. This event starts whena follicle reaches diameter of 10mm and more. FSH and sometimes gonadotrophin antagonists is required
because, for these patients, even if respond to natural events, their functioning follicles may, stop growing, orprematurely luteinised if left to themselves.
With this assumption, we started a protocol of delayed stimulation. These patients usually presented withraised FSH (>15U/L) on day 2-3, and typically AFC would be 0 or less than 2. No FSH is given as in routine
IVF then, but we would just do US every 3-5 days, depending on findings, until at least one follicle reaches adiameter of over 10mm, and then a low dose FSH of 75-150U sc per day is added. Antagonist is added whenfollicle diameter is 14mm or over, and egg collection when at least one follicle reaches 17mm. We presented
our preliminary data at the ISIVF in Montreal. We termed our treatment nurturing of these follicles tomaturity instead of ovarian stimulation.
This presentation deals with our collective data of over 200 cycles by two doctors working in the same clinicand doing IVF in the same IVF embryology laboratory. We will show that in over 95% of cases at least 1 egg
was collected which was fertilized. Most of these patients will then participate in our embryo banking service
where they will bank these single embryos until at least three, and then replaced as thawed-frozen transfer.
Overall, we have a clinical pregnancy rate of over 10% in this very difficult group of patients.
In conclusion we described a novel way of preparing the ovary for a group of patients who otherwise may not
be accepted into an IVF program. If they start IVF a lot ends up in cancelled cycles. We have shown that withpatience and care, a gentle "nurturing" of the ovary gives better result than super doses.

163

●Concurrent Symposium C-13.2◆Biosketch
Dr. Milton Leong has been involved in IVF since 1985. He received his medical education at McGill
University and after post graduate training also taught at McGill. In 1979 he returned to Hong Kong, and outof demand he started a practice in infertility. In 1985 he set up the IVF Centre, together with Dr. Clement
Leung, at the Hong Kong Sanatorium & Hospital and produced Hong Kong‘s first IVF baby in 1986.
In 2000 he was a leader to set up the Hong Kong Society for Reproductive Medicine, an interest he still
maintains. Dr. Leong is still in active practice and contributes to the specialty in publication and lectures. He
is a frequent speaker in regional and international meetings with current interest in identifying the threeportals of IVF: eggs, embryos and endometrium; and in poor responders and older patients.
Currently he is an adjunct professor of O & G of McGill University and a founder of the ivf-worldwide.com –the website for practicing IVF professionals.

164

●Concurrent Symposium C-13.3

Use of hCG in ovarian stimulation
Johan Smitz, MD PhD.
Research Director, Follicle Biology Laboratory (FOBI)
Faculty of Medicine and Pharmacy, Free University Brussels
Untimed LH increases are associated with reduced chances of ongoing pregnancy.
The etiologies of a high endogenous LH might be diverse: they may be part of WHO
type 2 anovulation or might be iatrogenically induced by increasing E2 levels (due to stimulation therapy).
Observing the relationship between a high basal or a premature rise of LH and reduced success-rates in ART
patients has led to the policy that LH should always be banned from the therapeutic arsenal of the
reproductive endocrinologist, with as only exception: the WHO-type I anovulation.
In 30 % of the patients having ovarian superovulation treatment using rFSH combined with GnRH agonist
therapy leads to an iatrogenic depletion of LH.
It was observed that making use of molecules with a longer halflife (hCG) in Highly Purified Human
Menopausal Gonadotrophins (HP-hMG) adds a significant qualitative element to treatment outcome (Diedrichet al., 2001; Andersen et al., 2006; Platteau et al., 2006, 2008). HCG concentrations in patient's serum (afterusing HP-HMG) on stimulation day 6 or day of the ovulatory stimulus was significantly positively correlatedwith the occurrence of a higher quantity of top quality embryos, higher pregnancy- and life birth rates.
Statistical modeling demonstrated that circulating hCG levels – independent from LH baseline concentrations– positively impact on the pregnancy outcome (Arce and Smitz, 2011). The precise molecular backgroundbehind these observations is now under further research.
Besides the differences in clearance there might be differences at the level of molecular interaction between
LH and hCG and the LH-receptor (LHR). The way that both molecules (LH or hCG) bind to the receptor
might induce a different cascade of molecular events downstream the receptor and/or lead to differences inreceptor metabolisation. It is known that a mutation in exon 10 of the LH receptor precludes effects of natural
LH, but in contrast hCG remains effective in stimulating the LH receptor. Mechanisms driving the superior
results by hCG over LH need to be investigated in relevant physiological models such as follicle cells andendometrial tissue.
◆Biosketch
Professor Johan Smitz (MD, PhD) is currently leading a research laboratory on ovarian and oocyte biology at
the Free University Brussels (VUB) and is Head of the Hormone Laboratory of the University Hospital VUB,
named UZBrussel. His clinical activity involves endocrine follow-up of infertility patients at the Center For
Reproductive Medicine at the UZBrussel.
His interests are in Oocyte Development in Vitro, Reproductive Physiology, culture systems and Clinical
Biochemistry.

165

●Concurrent Symposium C-13.4

Follicle stimulation by transdermal application of FSH
Herbert Zech, M.D., Professor
IVF Center Prof. Zech - Bregenz
This report establishes for the first time the effectiveness of transdermal delivery ofmolecules as large as follicle stimulating hormone (FSH, 32 KDa protein).
We chose IVF as a model to test the transdermal device because a clinical response
can be seen within days (by analysis of follicle growth) and the pregnancies testify to the quality of theretrieved oocytes. In the presentation, broader implications for the use of such a transdermal device (e.g.: thetransport of other peptides, proteins and vaccines of large molecular size or of hydrophilic nature across theskin) will also be discussed.
◆Biosketch
President of the Austrian Society for Reproductive Medicine and Endocrinology 2003-2006. Since 2007
President of Honor of the Austrian Society for Reproductive Medicine and Endocrinology.
After medical school at the University of Innsbruck, Herbert moved to the University of Graz and Innsbruck
for a professional training as a gynaecologist finishing this specialty in 1982. Herbert was always researchoriented and moved to the United States for a post doc at the University of Louisville Kentucky working on
basic cancer research with steroid receptors and vitamin A-receptors in rodents and in man. From there hewent back to the University of Innsbruck and became Head of the Department of Endocrinology. In 1984 hemoved to Bregenz and started the first IVF-Clinic realizing the big need for infertility treatment. The first IVF
children in southern Germany, Switzerland and western Austria were the results of the initial months oftherapy in Bregenz. In order to further develop this technique and the potential for the future he founded thefirst private clinic for in vitro fertilization in Austria and was also the first to be ISO certified worldwide in thefield of reproductive technologies. Realizing the deep desire for children, Herbert founded further IVF-centersin Italy, in Switzerland, in Liechtenstein, in the Czech Republic and in Salzburg, Austria. The total workforce
in all centers is 120 and is increasing. In recognition of his ongoing research efforts, Herbert was promoted
professor at the University of Innsbruck in 1999, where he teaches reproductive technologies andendocrinology. He is Harvard Business School Alumnus (Owner-President-Management Program – OPM39)
since 2010. Herbert brings into the company excellent knowledge of a broad spectrum in endocrinology,
genetics, embryology and stem cell research, a global network of medical collaborators, as well as provenentrepreneurial commercial business development experience.
IVF Center Prof. Zech - Bregenz, Römerstr. 2, 6900 Bregenz, Autria

166

●Concurrent Symposium C-14.2

Quality evaluation of human & mouse IVM embryo and epigenetic
evaluation of human born IVM babies
Hiroaki Yoshida
Yoshida Ladies Clinic, Center for Reproductive Medicine, Miyagi, Japan
Introduction: IVM oocyte exhibit lower viability and developmental potential after
fertilization. We compared the oxygen consumption rate of human IVM and COH
embryo with SECM non-invasively. Mitochondria regulate metabolism and influence the developmental
potential of oocytes and embryos. We evaluated basic research to clarify for IVM culture problem due tomitochondrial function using mouse IVM oocytes, and also evaluated chromosomal numbers and prematurechromosomal separation (PCS) with control and different medium (HTF & Waymouth). There was the reportof imprinting disorder with ART patients recently. So we checked the epigenetic disorders for born IVM andnatural pregnant babies.
Material and method: The oxygen consumption rate was calculated with SECM system for human oocytes,
IVM and COH embryos. We evaluated the number of oocytes, maturation rate and fertilization rate in human
IVM cases. And we compared the imprinting genes from umbilical cord of IVM and natural pregnant babies.
Basic research was performed using mice GV oocytes. We used different culture medium for oocyte
maturation and MII oocytes have done IVM/IVF, after that we evaluated embryo developments and theseveral mitochondrial function, and also evaluated, chromosomal number count, and PCS.
Results: The oxygen consumption rate for a single oocyte was as follows: GV 0.49, MI 0.47, MII 0.41 F ×
1014/mols-1. There were no significant difference in the mean oxygen consumption rate at each embryo inboth IVM & COH-IVF embryos (0.26-0.56 × 1014/mols-1). There were mean number of oocyte (10.4±5.3), maturation rate (54.7±18.7) and fertilization rate (85.8±15.4) in delivered cases. There was no
significant difference of epigenetic finding in all human IVM and natural pregnant babies. In the basic
research, HTF group was observed mitochondrial aggregation from like a spot in mitochondrial staining in
mice. There was lower MT membrane potentials in the HTF group than in vivo and Waymouth group.
Although there was no significant difference in total chromosomal count, PCS has much increased inWaymouth with small layer group.
Conclusion: There was no significant difference both IVM & COH embryo oxygen consumption rate after
fertilization. This means IVM embryo has same mitochondrial activity with COH-IVF embryo. The
mitochondrial function and chromosomal count & PCS were influenced from different IVM culture media.
Further study of cytoplasmic maturation and mitochondrial function from IVM is required.
◆Biosketch
1988 Assistant Professor : Division of Reproduction Dept. of OB/GYN Tohoku Univ.
1991 After acquired Ph.D. MIAMI School of Medicine Division of Reproductive Medicine Dept. OB/GYN
Director of IVF Lab.
1998 Opened Yoshida Ladies Clinic
2005 Division Chief of Center for Reproductive Medicine
2008 Clinical Associate Professor of Tohoku University Dept. of OB/GYN

167

●Concurrent Symposium C-14.3

Lessons from the mouse follicle culture and in vitro maturation models
Johan Smitz, MD PhD.
Research Director, Follicle Biology Laboratory (FOBI)
Faculty of Medicine and Pharmacy, Free University Brussels
Primordial follicles from juvenile mice ovaries can be grown in a petri dish during 3
weeks and produce oocytes capable being fertilised by sperm. Good health of theoffspring has now been confirmed on a large series of animals by John Eppig's Laboratory. Making theIn-vitro culture technology work for a broad range of follicle classes requires a whole range of different
culture techniques.
An unresolved question is how primordial follicles could be induced to grow in an ordered manner, instead ofactivating massively once they feel in vitro. Following the initial work from A. Hsueh's laboratory who could
induce invitro activation of primordial follicles in mouse and human ovarian tissue -which was
xeno-transplanted- and developed normally, our research addresses entire in-vitro conditions which could
induce controlled growth of prinordial follicles in ovarian tissue.
Work from several teams (Newton et al. (1995) in Sheep, Xu et al. (2009a) in Rhesus monkey , Xu et al.
(2009b) in human) have been able to grow secondary ovarian follicles in-vitro up to meiotic competence. Bymaking use of an extracellular matrix (matrigel, alginate) in a basal medium which maintains granulosa celldifferentiation (i.e. estrogenic) it is possible to grow these follicles and obtain oocytes with a capacity that is
sufficient to be fertilised. However at any time of the cycle there are only a limited number of such secondary
follicles present.
The fear that a prolonged culture period could change the imprinting pattern in oocytes during theit growthphase has proven to be unjustified (Anckaert et al., 2009).
As our knowledge advances, the chances of having one day an artificial ovary that could support full follicular
growth becomes more realistic.
◆Biosketch
Professor Johan Smitz (MD, PhD) is currently leading a research laboratory on ovarian and oocyte biology at
the Free University Brussels (VUB) and is Head of the Hormone Laboratory of the University Hospital VUB,
named UZBrussel. His clinical activity involves endocrine follow-up of infertility patients at the Center For
Reproductive Medicine at the UZBrussel.
His interests are in Oocyte Development in Vitro, Reproductive Physiology, culture systems and Clinical
Biochemistry.

168

●Concurrent Symposium C-14.4

Laboratory aspect of natural cycle IVF/M treatment
Ri-Chen Chian
McGill University, Montreal, Canada
Currently most infertility treatments employ ovulation stimulation with gonadotropins
to increase the number of oocytes available for in vitro fertilization (IVF), because it
has been believed that the success rate of IVF treatment relates directly to the number of embryos available
for transfer. Although the side effects of ovarian stimulation with gonadotropins, including ovarian
hyperstimulation syndrome (OHSS), have been well documented, the long-term side effects of repeatedovarian stimulation with protocols involving a gonadotropin-releasing hormone (GnRH) agonist or antagonist
in combination with gonadotropins are largely unknown. Immature oocyte retrieval followed by in vitro
maturation (IVM) of these oocytes is an attractive infertility treatment for women with infertility. In
comparison with ovary stimulated IVF treatment, the major advantages of IVM treatment include avoidanceof the risk of OHSS, reduced cost, and simplified treatment. Immature oocyte retrieval followed by IVM ofthese oocytes was initially shown to be a successful treatment for infertile women with polycystic ovary
syndrome (PCOS), because there are numerous antral follicles within the ovaries in this group of patients.
Immature oocyte retrieval followed by IVM might be useful in up to approximately 30% of women
undergoing IVF treatment who have large numbers of antral follicles including patients with PCOS. In general,clinical pregnancy and implantation rates of IVM treatment have reached approximately 35% and 15%,
respectively. Attempts have been made using IVM technology to treat patients with normal menstrual cycle
and intact ovaries. As protocols, the patients were given 10,000 IU human chorionic gonadotropin (HCG) 36 hprior oocyte collection in natural cycle when the leading follicle reached to 12-14 mm in diameter. The results
indicate that mature oocytes can be retrieved when the leading follicles reached 12-14 mm in diameter after
HCG administration. It is important to identify the mature oocytes at egg collection. This new technology has
been defined as natural cycle IVF combined with IVM, namely Natural cycle IVF/M, in order to distinguishwith traditional concept of IVM treatment. In this lecture, we will discuss laboratory aspects of natural cycle
IVF/M treatment in term of 1) Preparation of IVM media; 2) Identification of immature and mature oocytes;
3) In vitro maturation of immature oocytes; 4) Insemination of in vitro matured oocytes; 5) Embryo transfer.
◆Biosketch
Currently, Dr. Chian is Professor (rank as Associate Professor with Tenure) at Division of Reproductive
Biology, Department of Obstetrics and Gynecology, McGill University, Montreal, Canada. Dr. Chian is the
key person developed in vitro maturation (IVM) of human oocytes for clinical application and responsible forcryopreservation of human oocytes produced pregnancies and live births at McGill University Health Center(MUHC) in Canada. Dr. Chian published numerous research papers and presentations in refereed journals andinternational/regional conference. Dr. Chian edited two books, and contributed many book chapters. As aguest or invited speaker, Dr. Chian delivered numerous lectures in different societies and countries. Dr. Chian
was former Associate Editor for the journal, Human Reproduction and is an Editorial Board Member for
Journal of Assisted Reproduction and Genetics and other two journals. Dr. Chian is acting as a reviewer for
many other scientific journals, including Fertility and Sterility, Reproduction, Biology of Reproduction,
Reproductive BioMedicine Online, Endocrinology, The Journal of Clinical Endocrinology & Metabolism, and
The Lancet, etc. Dr. Chian‘s research interests are focused on 1) Ovarian function; 2) The mechanism ofoocyte maturation and activation; 3) Fertility cryopreservation. Dr. Chian speaks English, Chinese, Korean
and Japanese as well as some knowledge of French.

169

●Concurrent Symposium C-14.5

Cytoplasmic maturation and mitochondrial activity in human IVM
Yoshiharu Morimoto
IVF Namba Clinic, Osaka, JapanThe treatment for patient of polycystic ovary syndrome (PCO) may be the mostimportant by an increase of patients. At the time male factor infertility was almost
solved, the PCO is one of remained diseases that is difficult to manage. In order to achieve ovulation in thosepatients, it needs ovarian stimulation with more risk for ovarian hyperstimulation syndrome (OHSS). In vitro
maturation (IVM) of oocyte procedure would be a best option for PCO treatment in terms of complete
prevention of OHSS. Since the technology has been clinically applied, the number of centers undergoing it
has increased. Moreover it became to be applied for the normal menstrual cycle cases, poor quality embryocases and PGD source. However it cannot be said to be alternative for conventional IVF procedure, because
the clinical outcome has not been reached to the acceptable extent.
In IVM procedure, the immature oocytes in germinal vesicle or metaphase I stage are cultured with medium,
gonadotropins and serum for 24 to 48 hours. About 50 to 60 % of immature oocytes commonly maturate tometaphase II stage. In order to acquire better outcome, it is essential to elucidate maturation process of humanoocytes. Oocytes maturate in their nucleus and cytoplasm. The investigation has been concentrated to the
nuclear maturation process, but not for cytoplasmic maturation. In the cytoplasmic maturation, mitochondria
may have a key role for oocyte maturation and further development for embryogenesis in the organella.
They produce ATP as an energy source for resumption of meiosis, fertilization and cleavage. By ourultrastructural observation of oocytes in maturation process during IVM procedure, mitochondria haveshowed their unique appearances. In the germinal vesicle stage of unstimulated oocytes, mitochondria weredense and scattered in cytoplasm. In the second metaphase stage, mitochondria increased in number andaggregated in the center of cytoplasm.
We have investigated to express active mitochondria using porcine and human immature oocytes. Active
mitochondria were stained by DAB (3,3,-diaminobenzidine tetrahydrochloride ) on the ultrathin sections.
The appearance of mitochondrial distribution changes dynamically at each stage of maturation. We haveobserved it in JC-1 stained oocytes by confocal laser microscope.
In order to estimate viability of maturing oocyte, the cellular respiration during oocyte maturation with ascanning electrochemical microscopy was evaluated.
◆Biosketch
Prof. Yoshiharu Morimoto is currently Chief Executive Officer and Chairman of IVF JAPAN group which
includes two big infertility centers of IVF Namba Clinic and IVF Osaka Clinic running 6000 periods of IVF
cycles in total. He is also a visiting professor of Pochon CHA University, Saint Marianna University andKinki University Faculty of Biology-Oriented Science and Technology, and associate professor of Kyoto
University and Kansai Medical University. He was graduated from Kansai Medical University in 1977,
national board certified and obtained a degree of PhD in 1987. He is active in research area and his main
research field is ultrastructure in reproduction and concerns on oocyte maturation mechanism. He is mainly
focused on mitochondrial activity. Prof. Morimoto is the president of Japan Society of Assisted Reproduction,
former president of Japan Society of Fertilization and Implantation, and the president of the Asia PacificInitiative on Reproduction (ASPIRE).

170

●Concurrent Symposium C-14.6

Correct embryo selection improves embryo implantation rates in IVM
cyclesHai Ying Chen1, Alper Mumcu1, Seang Lin Tan1,2Montreal Reproductive Centre, Montreal, Canada1 and Department of
Obstetrics and Gynecology, McGill University, Montreal, Canada2
In vitro maturation (IVM) was introduced into clinical practice in the late 1980s. Since
then, it has become an increasingly important method in human ART. A successful IVM program provides
several advantages including avoidance of ovarian stimulation, reduction of treatment costs, and eliminationof the side effects of medication, including the risk of ovarian hyper-stimulation syndrome. Treatment
outcomes have improved substantially since we started our IVM program in 1998. In women younger than 35
years of age, our IVM clinical pregnancy rates have been comparable with average IVF clinical pregnancy
rates in North America. However, embryo implantation rates have been lower in IVM cycles and similar
clinical pregnancy rates have been achieved by transferring a higher number of embryos.
Between 1998 to June of 2003, we performed 279 IVM cycles reaching embryo transfer (ET). Mean female
age was 32.7 years, the mean number of embryos transferred was 3.4, resulting in a clinical pregnancy rate of
19.4%, with implantation rate of only 6.9%. Between 2003 July to 2010 December, we performed 502 IVM
cycles reaching ET. The mean female age was 32.8 years, and the mean number of embryos transferred was
3.5, resulting in a clinical pregnancy rate of 35.6%, with an implantation rate per embryo of 13.2%. Despite
the relative increase, figures were still low compared to IVF results. Since January 2011, contrary to theformer period when we essentially used to do cleavage stage embryo transfers, we adopted a policy ofpreferential blastocyst transfer for IVM patients. Based on 11 IVM cycles, with a mean female age of 31.4
years, and mean number of embryos transferred of 1.8, significantly less embryos transferred than in former
periods, we achieved a clinical pregnancy rate/cycle started of 36.4% with a mean embryo implantation rate of
31.8%.
Conclusion: Preferentially extending the culture period to five days and choosing the blastocyst with best
morphological characteristics seems to significantly increase embryo implantation rate. This also enables
elective single embryo transfer (SET) in IVM cycles with comparable clinical pregnancy rate with IVF as
achieved by most IVF centers. Multiple pregnancies can also be avoided with elective SET policy for IVM
patients.
◆Biosketch
Dr. Hai Ying Chen obtained his medical degree from Beijing University Medical School followed by aresidency in Beijing University First Hospital. He completed Master of Science degree at the Department ofExperimental Medicine in McGill University, Montreal, Canada. He continued as a research associate in the
Department of Obstetrics and Gynecology, McGill University. He worked as a clinical embryologist at
McGill Reproductive Center between 2008 and 2010. Dr. Chen is currently working as the director of in vitro
maturation and oocyte vitrification programs of the Montreal Reproductive Center. He has conducted several
research projects on IVM and oocyte vitrification. He has authored or co-authored several publications andhas been invited to lecture in various international conferences.

171

●Concurrent Symposium C-15.1

IVM Application in PCOS Patients
Xiaoying Zheng, Lina Wang, Ying Lian, Xiumei Zhen, Ping Liu,
*Jie Qiao
Reproductive Medical Centre, Peking University Third Hospital,
Beijing, P.R. China
Over the next decade, immature oocyte retrieval followed by IVM becomes a widespread treatment for
infertile women with PCOS because there are numerous antral follicles within the ovaries in this group ofpatients. Compared with ovary-stimulated in vitro fertilization (IVF) procedure, the major advantages of IVM
include avoidance of the risk of ovarian hyperstimulation syndrome, reduced cost, and simplified treatment.
Although good results have been reported by some clinics， in-vitro maturation has not yet become amainstream fertility treatment. The most important reason for this is the lower chance of a live birth per
treatment compared with conventional in-vitro fertilization.
Gonadotropin plays an important role in the regulation of oocyte growth and maturation. In order to mimic thepreovulatory luteinizing hormone (LH) surge in spontaneous menstrual cycle, human chorionic gonadotropin(hCG) as a surrogate for LH is administered at a dose of 5000-10000IU at the end of follicular stimulation to
trigger the meiosis resumption and nuclear maturation of oocytes in IVF technology. The traditional
applications of hCG have proven to be highly successful and valuable tools in the treatment of infertility forover four decades. However, the effect of hCG priming on oocyte maturation and developmental competencein IVM cycle has remained a debatable issue for several years. Chian and colleagues demonstrated that hCGpriming could hasten the maturation time of the oocytes in PCOS women. Subsequently, a multicentre study
by the same investigators have further supported this finding by reporting pregnancy rates of 30-35% in
hCG-treated IVM cycles in patients with PCO and PCOS. However, another studies did not demonstrate any
beneficial effect of HCG priming. Although there are many reports of pregnancy rates after IVM withgonadotropin priming, there is little data of clinical controlled randomized study. Therefore, the present
controlled randomized study was designed to determine whether hCG priming prior to oocyte aspiration can
improve embryonic developmental competence and clinical result generated from IVM oocytes of
unstimulated women with PCOS.
Objective: To investigate whether hCG priming prior to oocyte aspiration can improve embryonicdevelopmental competence and clinical result generated from IVM oocytes of unstimulated women with
PCOS.
Patients: 82 women with polycystic ovary syndrome (PCOS) undergoing 83 IVM treatment cycles.
Interventions: Each patient was randomly assigned either to be primed with 10000IU hCG or not primed
36-38 hours before oocyte retrieval. After the oocytes had matured in vitro, fertilization and embryo transfer
were performed.
Result: The average number of cumulus-oocyte complexs (COCs) recovered were 13.8 and 14.7 in thehCG-primed and non-primed groups respectively (p>0.05). The maturation rate of COCs was significantly
improved in the hCG-primed group (55.43% versus 42.56%; p<0.05). The fertilization and cleavage rates
were comparable between the two groups. There was no significant difference in the clinical pregnancy(37.50% vs 48.84%), live birth (22.50% vs 30.23%) and implantation rates (32.86% vs 31.46%) between

172

●Concurrent Symposium C-15.1

hCG-primed and non-primed group. The number of pregnancy loss were 6 (hCG-primed group) and8(non-primed group) cases respectively,
Conclusion: While a significant improvement of maturation rate of immature oocytes were observed in
hCG-primed IVM cycle with PCOS patients, the use of hCG prior to oocytes retrieval does not improve thesubsequent embryo developmental competence. Higher rate of pregnancy loss was observed in IVM cycle.
◆Biosketch
Qiao Jie is Chief Physician and head at the Department of Obstetrics and Gynecology of Peking University
Third Hospital and is now the president-elect for reproductive medical branch of Chinese medical association.
She went to Queen Mary Hosp, the University of Hong Kong as a visiting scholar from 1996 to 1997 andStanford University Medical Center as Postdoctoral Research Scholar from 2002 to 2003. She specializes inclinical treatment, education and medical researches on female fertility preservation and improvement, as wellas the diagnoses and treatment for infertility and reproductive endocrine diseases, the focus of PCOS. She
published over 200 articles in medical journals in the field of OB/GYN. Her researches have been fundedthrough variety of grants, includes National Outstanding Youth Fund and Key Technologies R&D Program.
She is Treasurer of Asia-Pacific Society for Reproductive Medicine, Board member of AEPCOS Society andAsian Journal of Andrology, Associate Chief Editor of Chinese Version for Fertility and Sterility, Editorial
Member for Reproductive Biology and Endocrinology and Seminar Reproductive Medicine. She has received
many awards including National Science and Technology Progress Award Ministry of Scientific ProgressAward first prize. Qiao becomes to be the head scientist of Major State Basic Research Development Programof China (973) in 2011.
Qiao Jie
Professor, M.D.
Department of Obstetrics and Gynecology,
Peking University Third Hospital, No.49 North Huayuan Road, Haidian District, Beijing, China, 100191

173

●Concurrent Symposium C-15.2

Oocyte maturation from tiny follicles in human ovarian tissues
Shu Hashimoto1, Yodo Sugishita2, Nao Suzuki2, Bunpei Ishizuka2,
Aisaku Fukuda3, Yoshiharu Morimoto1
1
IVF Namba Clinic, Osaka, Japan,
2
St. Marianna University School of Medicine, 3IVF Osaka Clinic
Ovarian tissue cryopreservation has been shown to be effective measure for fertility
preservation. However, this method is inadequate for freezing of antral follicles. To preserve Immature
oocytes in antral follicles, cryopreservation of IVM oocytes is a valid procedure after the culture of immature
oocytes retrieved following follicle aspiration or follicle isolation from ovarian tissue excised by laparoscopy.
Thus, the combination of in-vitro maturation (IVM) of immature oocytes and oocyte freezing is one of usefulmeasure to preserve the fertility of patients who undergo chemotherapy and/or radiotherapy, and are short oftime for controlled ovarian hyperstimulation (COH, Hashimoto et al., in press). After the approval of IRB of
St. Marianna University School of Medicine, we started the cryopreservation of mature oocytes and ovaries
for patients who were at risk of gonadal dysfunction from cytotoxic chemotherapy or radiotherapy. Thus far,
63 immature oocytes were obtained from 1- to 5-mm follicles of five patients, of which 32 were matured(51%). Thirteen mature oocytes were cryopreserved. The maturation rate was similar to that of oocytes
retrieved from approx. 10-mm follicles in routine IVM protocol in which immature oocytes are retrieved fromlarger follicles because of a technical challenge of low recovery rate from smaller follicles. IVM of immature
oocytes following their retrieval has been proposed as a potential alternative to conventional IVF treatment
following COH. However, low developmental competence of IVM oocytes compared with that of in vivo
matured oocytes prohibits the progress of IVM protocol. To overcome this challenge, several studies haveexamined the effects of holding oocytes at the germinal vesicle (GV) stage before IVM in cattle and mice,
because oocytes might require time to acquire developmental competence during meiotic arrest. This idea isbased on the fact that mammalian oocytes are arrested at the diplotene stage until the GnRH surge occurs andimmature oocytes resume meiotic maturation spontaneously following the release from follicles. Several
groups achieved the improvement of the developmental competence of IVM oocytes in cattle and mice
(Hashimoto et al., 2002; Albuz et al., 2010). An adaptation of these new culture systems for clinicalapplication might have significant implications for infertility management. Hashimoto et al., Reprod Med Biolin press. Hashimoto et al., Biol Reprod 2002; 66:1696-1701.Albuz et al., Hum Reprod 2010; 2: 2999-3011.
◆Biosketch
Dr Hashimoto obtained his PhD in Reproductive Physiology at Kyoto University in 2001. He started his
research career in 1989 by production of transgenic mice, rat, rabbits and cattle. He also developed assisted
reproduction technology (ART) in cattle (super ovulation, transvaginal oocyte retrieval, IVM, IVF, ICSI,
embryo culture and vitrification) in Snow Brand Milk Products Co., Ltd. He moved to IVF Namba Clinic in
2004 and started human ART. Currently, he is the research director of IVF Namba Clinic. He was recently
secretary-general of the Japan Society of Fertilization and Implantation. He received the JSAR (JapaneseSociety of Animal Reproduction) innovative technology award in 2008 and JSMOR (Japanese Society of
Mammalian Ova Research) outstanding presentation award in 2009.

174

●Concurrent Symposium C-15.3

Natural Cycle IVF combined with IVM
Jin-Ho LIM, MD
Maria Fertility Hospital
IVM has been developed to prevent the side effects of ovarian hyperstimulation, andthe efficiency and safety of IVM were already proved.
The clinical pregnancy rate of IVM reached 35-40%/ET, and more than 2000 healthy IVM babies have beenborn in the world.
But the main indication of IVM was PCOS patients with irregular anovulatory cycles.
To expand the indication of IVM for patients with regular ovulatory cycles, we designed the new procedure
what we like to call ' Natural cycle IVF combined with IVM (Natural IVF/M)'.
We check baseline ultrasound on MCD #2-3.
If the number of AFC is more than 7, we recommend Natural IVF/M for patients with regular cycles.
When the leading follicle reaches 12-14mm in diameter, we give 10,000IU of HCG, and collect the oocytes
36-38 hours later.
The matured oocytes collected at the time of OPU are inseminated by ICSI on the same day, and the immature
oocytes are cultured in vitro for 24-48 hours.
In vitro matured oocytes are then inseminated by ICSI subsequently.
On day 3 or 4 after OPU, the embryos are pooled together and the best 1-3 embryos are selected for transfer.
We could confirm that the Natural IVF/M together with IVM are an efficient treatment for more than 50% ofinfertile women with acceptable clinical pregnancy rate and we could do embryo transfer in almost every
cases.
◆Biosketch
Dr. Jin-Ho Lim is the founder of Maria Medical Group.
He is an internationally recognized infertility expert and a pioneer in simplifying IVF treatment. He
established the Maria Fertility Hospital in 1989 and produced the first natural cycle IVF baby in Asia.
In the end of 2006, more than 30,000 IVF babies had been born in his 9 IVF Centers in Korea, and they are
performing more than 10,000 OPU cycles per year for infertile couples.
He also has an IVF center in Beijing, China.
Maria IVF Centers have been offering IVM or Natural cycle IVF/IVM treatment for patients who has morethan 7-8 antral follicles at the beginning of menstruation.
In addition, Dr. Lim is the President of Maria Biotech. Institute in Seoul, Korea, where they first generated
human embryonic stem cell (HESC) lines in Korea, and the first 3 of them were registered by NIH, the UnitedStates.
Dr. Lim has published many original scientific papers and review articles, and delivered many lectures andpresentations in national and international conferences.
Dr. Lim‘s recent research interests include: 1) Simplification of IVF treatment; 2) Human placental stem cells.

175

●Concurrent Symposium C-15.4

The effective approach for IVM-IVF using Metformin
Aisaku Fukuda
The Centre for Reproductive Medicine and Infertility
Introduction: Although IVM-IVF (In vitro maturation, in vitro fertilization andembryo transfer) is a relatively new option for ART with significant benefits forpatients, its success rate is still less than conventional ART. First successful IVM-IVF in the world wasachieved in 1994, 16 years after first IVF. The present study was conducted to determine if Metformin
administration benefited on outcomes of IVM-IVF in PCO patients.
Materials and Methods: We have performed 914 cycles of IVM-IVF (571 fresh and 343 frozen cycles) andhave achieved 122 successful pregnancies (23%) from Oct. 1999 through Mar. 2011, since our first success inJapan. When we divide the last 12 years into two terms (term 1: 2000-2005 and term 2: 2006-2011) andcompare them, the pregnancy rates of term 2; total, fresh and frozen (33.8%, 35.6% and 30.6%) was
significantly higher (P<0.01) compared to term 1 (16.2%, 19.1% and 15.7%, respectively). Multiple strategies
such as double needle system, frozen cycles, low dose FSH, various culture media, HCG pretreatment andMetformin, alone or in combination have been attempted in the meantime. We focused on Metformin in thepresent study, because Metformin was routinely used last 6 years. Fresh cycle IVM-IVF was performed on
172 PCO patients in term 2, either with Metformin of 1500mg/day for at least four weeks (Group A; n=74) orwithout Metformin (Group B; n=98). Follicular monitoring began from day 7, and if needed, 150 units/day of
FSH were administered until follicles reached 10 mm in diameter. HCG was administered 36 hours before
retrieval. Immature oocytes were cultured in IVM medium (MediCult) supplemented with 10% SSS (Irvine)for 26 hours, and ICSI was performed on in vitro-matured oocytes. Fertilization was confirmed 18 hours after
ICSI. Day 3 embryos were transferred after assisted hatching.
Results: The pregnancy rate, number of oocytes retrieved and good quality embryos in group A (51.1%, 12.1
and 2.1) were significantly higher (P<0.01) than in group B (29.6%, 8.4 and 1.0, respectively). However, there
were no significant differences in the rates of maturation (53.7% vs. 51.9%), fertilization (81.9% vs. 83.4%),
and good quality embryos (36.2% vs. 29.3%).
Conclusions: The present study suggests that Metformin pretreatment of PCO patients improves clinicaloutcome of IVM-IVF by increasing the number of oocytes retrieved and good quality embryos by altering
intrafollicular environment. IVM-IVF might be applied as a first choice of ART for PCO patients with
Metformin pretreatment.
◆Biosketch
Aisaku Fukuda, M.D., Ph.D., HCLD (ABB) Chairman; IVF Osaka Clinic, Clinical Prof.; Kansai Medical
University, Osaka Japan, Clinical Prof.; East Tennessee State University, USA
1978: M.D. Kansai Medical University, Osaka Japan
1978-1980: Residency Kyoto University, Kyoto Japan
1980-1984: Director, Maizuru City Hospital, Dept. OB/GYN. Kyoto Japan
1984-1988: Graduate school, Kyoto University Faculty of Medicine, Japan
1989: Ph.D. Kyoto University
1990-1992: Research associate, East Tennessee State University (ETSU), USA
1996: High Complexity Laboratory Director (HCLD) by American Board of Bioanalyst (ABB)
1993-1998: Assistant professor, (ETSU), USA
1998-present: IVF Osaka Clinic, Osaka Japan
Awards: AFS (ASRM) Poster Prize Award 2nd place (1992),Kansai Medical University Alumni Association
Award (1993), Sigma scientific photography prize (1995), Japan Society of Fertilization and Implantation
Award (2000), Good speech Award from International Laser Medicine Congress (2009)176

●Concurrent Symposium C-15.5

Clinical implications of IVM
Svend Lindenberg Professor dr. med.,Mette Munk, Chief Embryologist
Copenhagen Fertility Center, Copenhagen
With the recent update of IVM children born (Ri-Cheng Chian), it is obvious, that
IVM has a place in routine ART. Indeed also in combination with the newly reported
excellent results following vitrification of oocytes.
Three different aspect of IVM are today applicable to routine IVF:
1) IVM of immature oocytes from single women wanting to preserve fertility by freezing oocytes
2) IVM as backup to IVF in high responder to reduce the OHSS risk
3) IVM/IVF in routine ART as the efficient treatment option for all IVF. Specifically to allocate women in real
IVM, real IVF and IVM/IIVF cycles.
By applying this approach in the IVF treatment several goals are archived:
1) Reducing the medication
2) Reducing the risk for OHSS syndrome◆Biosketch
Professor dr. med Svend Lindenberg is a pioneer in IVF and has partcipated in the group of scientist
producing the first IVF baby in Denmark in 1982. Since then published more than 150 papers and given
numerous presentations in internatinal and national societies and holds several international patents in ART.
Today director for Copenhagen Fertility Center and Research Institution with focus on low stimualtion and
IVM.
Copenhagen Fertility Center: Copenhagen, Lygten 2c, DK-240 NV

177

●Concurrent Symposium C-16.2

Early embryo development: What’s criticalMoncef Benkhalifa1,2, Paul Cohen Bacrie2, Alain dalleac2,
Yves Menezo3
1
ATL R&D. Reproductive Biology & Genetics Laboratory. Paris
2
Eylau Laboratory/ Unilabs. Paris-Geneva
3
Dynabio Laboratories. Cherbourg France
In functional biology, the fertilisation success is depending on oocyte maturation at genomic and epigenetic
level, in parallel to sperm gemone integrity. Sperm-oocyte fertilization and activation is a complicatedfascinated phenomenon in human reproduction and its molecular elucidation is still totally unknown becauseof the limitations of the material and difficulties to analyse the complicate proteins-proteins interactions.
Parallelly the oocyte surface is covered with microvilli with the exception of the region overlying the meiotic
spindle (Kaji et al 2004). Recent studies showed that genes targeting is a powerful tool to identify the functionof candidate molecules involved in sperm oocyte fusion and activation (Cuasnico et al 2001). Ubiquitous
tetraspanin family including CD9 are involved in sperm-fusion and for multi-molecular complexes withthese associated molecules on the plasma membrane, and thus have been implicated in cell adhesion, motility,
proliferation and differentiation (Boucheix et al 2001).
Participation of oocyte-specific GPI-anchored proteins in the sperm-oocyte interaction was suggested andoocyte specific phosphatidylinositol glycan class A (PIGA) deficient showed severely reduced fusibility ofoocyte with sperm (Alfeiri et al 2003). Also intention had been given to integrins on oocytes and their ligands
as members of ADAM family. After the interaction between gametes to activate the oocyte, the G protein
receptor is activated by the sperm binding proteins which activate tyrosine kinase which then activate PLC.
The inositol system is implicated as the pathway in egg activation via the phospholipase C (PLC).
It‘s possible also that soluble sperm factors diffuses from the sperm into the egg cytosol upon sperm oocyte
fusion by activating a signal transduction pathway that uses second messengers. A new PLC isoform, PLC
zeta may be equivalent of the mammalian sperm factors. Oocyte activation after fertilization go via a complex
pathway of genomic et epigenics maturation for maternal RNA reservoir to follow early zygote formation andembryo development until 4 cells, the stages of embryo genome expression◆Biosketch
Dr. Moncef Benkhalifa obtained his Ph.D. in 1992 from Clermont Ferrand University. He is qualified in
Reproductive Biology and Medical Cytogenetics from the school of Medicine and mainly involved in ART
and Genetics practice. Since 2006 he is consultant in reproductive biology & genetics at Eylau laboratories /Unilabs Paris and technical director of Unilabs, France.
Dr Benkhalifa is collaborating with different teams involved in ART and Genetics for clinical practice andresearch. He is author and co-author of more than 80 national and international publications with a maininterest for R&D in Embryology and Genetics.

178

●Concurrent Symposium C-16.3

Implantation window
Atsushi Azumaguchi
KKR Sapporo Medical Center Tonan Hospital
1. What is an implantation window? By resecting the ovaries of a female rat aftermating and continuously administering progesterone to the rat, a neutral state can becreated in which the blastocyst can survive in the uterus, but cannot implant. When estrogen is administeredduring this neutral state, a receptive state for the blastocyst appears within 24 h and lasts only 12 h. This 12-h
receptive state is referred to as the implantation window, and the blastocyst cannot implant after the end of theimplantation window (state of non-receptivity). Establishment of an implantation window in humans has not
yet been achieved. However, in histological studies of uteri following total hysterectomy, blastocysts wereobserved floating in the uterine cavity, but no implantation was confirmed up to day 19 of the estimatedmenstrual cycle and blastocyst implantation occurred only on day 21 or later, suggesting that an implantation
window similar to that in rats exists in humans.
2. Are pinopodes an index of implantation window? Since 1971, pinopodes have been considered to represent
an index of the implantation window. However, pinopodes have recently been claimed to be observed before
ovulation as well as during the luteal phase and pregnancy. If such observations are correct, pinopodes do not
represent a suitable index. We speculate that the above claim has been made because no detailed definition of
pinopodes has been established. Regarding pinopodes during pregnancies, we have also confirmed theexistence of pinopodes in the endometrium of tubal pregnancy. However, one possibility could be thatpinopodes disappear when pregnancy is not established and continue to exist when pregnancy is maintained.
Therefore, in our view, pinopodes can be used as an index of the implantation window.
3. Pinopodes in the natural cycle, human menopausal gonadotropin (HMG) cycle and hormone replacement
cycle: Pinopodes have been reported to be triggered by progesterone. In the HMG cycle, the amount ofpremature secretion of progesterone is larger and pinopodes have been suggested to appear earlier comparedwith natural cycle. In the hormone replacement cycle, pinopodes have been suggested to appear later than in
the natural cycle, due to a lack of progesterone secretion. We have obtained similar findings. In Japan, frozen
embryo transfer has resulted in a higher pregnancy rate than fresh embryo transfer, suggesting the possibility
that pinopodes developing early in the HMG cycle contribute to low pregnancy rates.
4. Pinopodes in abnormally thin endometrium: We found that pinopodes appear significantly earlier inabnormally thin endometrium less than or eqaul to 6 mm in thickness during the implantation phase than innormal endometrium more than or equal to 7 mm. We also found that the surface of the endometrial
epithelium was deteriorated with abnormally thin endothelium. We therefore speculated that early
development of pinopodes and deterioration of endometrial epithelium are factors for the low implantation
rate with abnormally thin endometrium.
◆Biosketch
Name: Atsushi Azumaguchi
Nationality: Japanese
Education: Medical school, Sapporo Medical University Post Graduate School, Sapporo Medical University
Major Research: Reproductive Endocrinology, hysteroscopic surgeryPresent
Position: KKR Sapporo Medical Center Tonan Hospital, Dept. of Reproductive Endocrinology

179

●Concurrent Symposium C-16.4

Luteal phase support
Svend Lindenberg Professor dr. med.
Copenhagen Fertility Center, Copenhagen, Lygten 2c, DK-240 NV,
It is obvious that any stimulation regime for IVF involving a high Estradiol andprogesterone serum level in the patient at the peri ovulatoric phase involves corpus
luteum malfunction. To rescue this several treatments protocols has been devised
such as:1) Low dose hCG
2) Vaginal progesterone
3) Low dose antagonist
Today the intramuscular and the vaginal supplementation are mostly used due to the risk of OHSS by using
hCG. The most interesting findings during the last year is that no patients need more than 16 days ofprogesterone supplementation following routine IVF treatment according to new published data. Further dataon low doses hCG as supplementation might again be a convenient part of the IVF treatment in the luteal
phase.
◆Biosketch
Professor dr. med Svend Lindenberg is a pioneer in IVF and has partcipated in the group of scientist
producing the first IVF baby in Denmark in 1982. Since then published more than 150 papers and given
numerous presentations in internatinal and national societies and holds several international patents in ART.
Today director for Copenhagen Fertility Center and Research Institution with focus on low stimualtion and
IVM.
Copenhagen Fertility Center: Copenhagen, Lygten 2c, DK-240 NV
In 2012 Congres President for the 2012 internationa Meeting in ISMAAR: www.ismaar2012.dk

180

●Concurrent Symposium C-16.5

Predictors of IVF outcome with a particular interest in the amount ofgonadotropin administeredPeter Kovacs, Steven G Kaali
Kaali Institute IVF Center, Budapest, Hungary
Background: Recently interest was renewed in the benefits of using less gonadotropin(G) for IVF. Mild stimulation is patient friendly and has economic advantages but theliterature is split on its efficacy. Our aim was to assess predictors of IVF outcome with a particular interest in
G dose.
Methods: All fresh, non-donor cycles that progressed to embryo transfer in 2009 were considered for the
analysis (N=1385). Cycles using GnRH agonist/ antagonist protocols were included. Patient (age, CD3 FSH,
order of treatment cycle, indication) and stimulation (protocol, G dose, # of follicles/eggs, embryo quality, #of embryos transferred, cryopreservation, day of transfer) were compared between cycles with and withoutclinical pregnancy (CP). The impact of medication dose was compared in cycles where below and above
median dose was used and across tertiles and quartiles of medication dose. Univariate & multivariable logistic
regression analyses were used.
Results: The overall pregnancy rate (PR) was 32.9%. In those cycles that resulted in a pregnancy the CD3
FSH was lower, the # of eggs and the proportion of good quality embryos was higher. Blastocyst stagetransfer was associated with a 58% improved chance for CP. The availability of surplus embryos forcryopreservation was associated with better outcome. The use of the GnRH agonist long protocol was more
likely to result in a CP when compared to the flare or antagonist protocol and the diagnosis of tubal infertility
was associated with lower pregnancy rates. According to our analysis the number of embryos transferred andendometrial thickness were not associated with CP. In those cycles that resulted in a pregnancy the mean daily
medication dose administered was less (175 IU +-59sd vs 195 IU +-73sd; p"<"0.00001). When comparisonswere made across dose tertiles pregnancy rates (36.8% vs 34.6% vs 26.8%; p=0.004) and implantation rates(20.5% vs 19.2% vs 13.8%, p=0.003) decreased as higher amount of G was used. Logistic regression analysis
showed that every additional ampoule of G daily decreased the chance of achieving a pregnancy by 17% (OR:0.83, 95% CI: 0.70-0.98, p=0.03).
Conclusions: Our results confirm findings of previous reports of predictors of CP. In addition we found thatthe use of less G is associated with higher pregnancy rates. These results lend support to the use of mild
stimulation but the ideal candidate and clinical setting for this still needs to be clarified.
◆Biosketch
Short bio for Peter Kovacs
I graduated from the Albert Szent-Gyorgyi School of Medicine in Szeged, Hungary in 1994. Upon graduation
I decided to follow my grandfather‘s and father‘s footsteps and chose to become an obstetrician gynecologist.
I did my ObGyn and Reproductive Endocrinology and Infertility training at the Albert Einstein School of
Medicine in New York. Upon completion of residency and fellowship training I moved back to Hungary to
my country of origin where I currently serve as the medical director of the largest IVF Center, the Kaali Clinic
in Budapest. I earned a PhD degree for studies assessing the effect of diabetes/insulin on GnRH neuron
activity using a rat model. My current research interest focuses on stimulation options for IVF, predictors of
IVF outcome and on various tools that could be used to improve laboratory and clinical outcome. Besides
work my 4 sons keep me busy and have a long standing love for tennis that I try to play when time allows.

181

●Concurrent Symposium C-17.1

Whole ooplasmic replacement in Germinal Vesicle oocytes
Akiko Yabuuchi
Advanced medical research institute of fertility, Kato Ladies Clinic
Oocytes from aged individuals have been known to have higher rates of aneuploidy
and higher proportions of mutated mtDNA compared to those from younger
individuals. Aneuploidy has been reported to be one of the major causes of infertility in older women. It has
been proposed that cytoplasmic factors in oocytes are responsible for the construction of meiotic spindle andindeed, dysfunctional cytoplasmic factors which have been shown in some aged oocytes increase thefrequencies of abnormalities in spindle formation and chromosome segregation. Therefore, theseabnormalities may be avoided by replacement of compromised cytoplasm with healthy cytoplasm through GV
transfer (GVT) before the start of chromosome segregation. This strategy can also be adopted for
mitochondrial disease patients by transferring patients' GV into enucleated GV stage oocytes containing
normal mtDNA. However, currently available methods for GVT is cell fusion method which karyoplast isfused with enucleated oocyte by inactivated Sendai virus (HVJ) or electric pluses. Karyoplast contains nucleus
and cytoplasm (mitochondria), therefore, the resulting reconstructed oocytes have mixture of two different
mtDNA in the cytoplasm which so called mtDNA heteroplasmy and the heteroplasmy persists if mtDNA fromkaryoplast actively replicate throughout embryo development and possibly will transmit in germ cells, as
determined by mitochondrial bottleneck. Therefore, the best way to reduce the risk of heteroplasmy is toremove patient mtDNA completely when GVT are performed. As an alternative to cell fusion methods which
results in various degrees of mtDNA heteroplasmy, we have established a new method in which we inject GV
without any visible residual ooplasm into enucleated GV stage oocyte using a Piezo-driven system (Germinal
Vesicle injection: GVI). We used a bovine model to explore the feasibility of GVI, as the size of GV stage
bovine oocyte is similar to that of human. Germinal vesicles were aspirated from the oocytes, cytoplasm
surrounding GVs were removed by pipetting and directly injected into enucleated GV stage oocytes. In ourstudy, the frequencies of in vitro maturation and blastocysyt development after fertilization of GVI oocytes
were 81% and 22%, respectively. These developmental endpoints were comparable to those ofnon-manipulated control GV stage oocytes (maturation: 81%, and blastocyts development: 25%). After
staining GVs with MitoTracker CMXRos, we confirmed that barely detectable amounts of mitochondria wereattached to GV membrane. Preliminary results of our study suggest that GVI is very effective method in orderto reconstruct GV stage oocytes without compromising their ability to complete meiosis. Although we believethat the residual cytoplasm transferred in to the reconstructed oocyte through GVI is extremely small, actual
fate of these mitochondria in developing embryos and offspring needs to be further evaluated in order to bring
the promise of clinical application.
◆Biosketch
2003.3 PhD at Laboratory of Animal Reproduction, Graduate School of Kinki University, Nara, Japan
2003.3-2004.12 Postodoctoral Research Fellow at Cellular Reprogramming Laboratory, Michigan State
University, East Lansing, MI, USA
2005.1-2009.10 Staff Scientist at Department of Hematology and oncology, Children's Hospital Boston,
Harvard Medical School, Boston, MA, USA
2009.11-2011.5 Scientist at Advanced Medical Research Institute of Fertility, Kato Ladies Clinic, Tokyo,
Japan
2011.6-Present Senior Scientist at Advanced Medical Research Institute of Fertility, Kato Ladies Clinic,
Tokyo, Japan182

●Concurrent Symposium C-17.2

Dynamic genome dysfunction and chromosomes abnormalities
Moncef Benkhalifa1,2, Aygul Demirol3, Alain dalleac2, Timur Gurgan3
1
ATL R&D. Reproductive Biology & Genetics Laboratory. Paris
2
Eylau Laboratory/ Unilabs. Paris-Geneva
3
Genetics & IVF, Gurgan Clinic, Ankara, Turkey
The contribution of chromosomes abnormalities, genes disorders and functional biology process miss fit inreproductive failure are well documented. The pre zygotic gametes maturation and competency are mandatoryfor early embryo cleavage, and development. More than the culture conditions, genetic and epigenetic statusinvestigation of the early embryo are important to select and predict the chance of clinical pregnancy in ART
programme. To select and predict the best gametes and embryo, Cytological, Cytogenetics, Moleculartechniques (karyotype, FISH, PCR and sequencing) or imaging methods are routinely applied sine many years
at pre and post zygotic stages to assess gametes and embryos quality. Recently, Genomic or expression
microarray technology becomes a common tool for genome and gene investigation in research anddevelopment and clinical diagnostic. For genome profiling and high resolution molecular karyotyping, arraycomparative genome hybridisation (array CGH) methods appear to be far better and have much higher
sensitivity and specificity for subtle genomic changes. In Reproductive Medicine and IVF, nanotechnology(genomic, expression or proteins arrays) is becoming an important clinical assay for genomics,
transcriptomics and proteomics testing. For non invasive testing cumulus cells, endometrial cells
transcriptome profiling, the metabolic monitoring of culture media can give valuable information on embryo
implantation competency and IVF success rate improvement.
◆Biosketch
Dr Moncef Benkhalifa obtained his Ph.D. in 1992 from Clermont Ferrand University. He is qualified in
Reproductive Biology and Medical Cytogenetics from the school of Medicine and mainly involved in ART
and Genetics practice. Since 2006 he is consultant in reproductive biology & genetics at Eylau laboratories /Unilabs Paris and technical director of Unilabs, France.
Dr Benkhalifa is collaborating with different teams involved in ART and Genetics for clinical practice andresearch. He is author and co-author of more than 80 national and international publications with a maininterest for R&D in Embryology and Genetics.

183

●Concurrent Symposium C-17.3

A novel trial of nuclear transfer for repairing an aged oocyte
Atsushi Tanaka1, Motoi Nagayoshi1, Masataka Yamamoto1,
Izumi Tanaka1, Hiroshi Kusunoki2, Seiji Watanabe3
1Saint Mother Hospital, Fukuoka, Japan,
2
Faunal Diversity Sciences, Graduate School of Agriculture, Kobe
University, Japan,
3
Department of Anatomy, Hirosaki University School of Medicine, JapanThe main causes of repeated failure in assisted reproduction such as IVF-embryo transfer are believed to beooplasmic deficiencies, abnormalities and ageing rather than nuclear deficiencies. It is a common
phenomenon that pregnancy rates decrease, but miscarriages increase, as women grow older. Also, thepercentage of fetal chromosomal abnormalities in miscarriages increases according to female age, reaching>90% when women are over 40 years old; surprisingly, about 90% of them are cases of autosomal trisomy.
Such aneuploidy is mainly induced by the chromosomal pre-division, in which homologous chromo-somes
fail to pair during meiosis I and segregate before it is complete, resulting in disomic gametes. Nuclear transfer
into the metaphase-II (M-II) oocytes shows promise as a means of repairing female infertility due toooplasmic deficiency and abnormalities. We therefore conducted nuclear transfer of in vitro matured
metaphase-II oocytes (recipient oocytes) into enucleated freshly ovulated metaphase-II oocytes (donor
oocyte).
In both in-vitro matured oocytes and freshly ovulated oocytes, the M-II chromosomes were easily recognized
as a round transparent substance in which the chromosome body was centrally located, and they were usually
beneath or adjacent to the 1PB with the aid of an inverted microscope equipped with a Normarski differential
interference contrast system. The aspirated M-II karyoplast of recipient oocytes was transferred into theperivitelline space of an enucleated donor oocyte. The grafted oocyte was transferred in Zimmerman cellfusion medium. Membrane fusion was facilitated by electrical stimulation (10V for 1 second AC + 10V for 45
microsecond DC) with an electro cell fusion generater (LF 201). After fusion, the constructed oocytes were
cultured in HTF medium for 2 hours and ICSI was performed.
The percentage of identification of M-II chromosome was 91.1 % (41 out of 45) in freshly ovulated oocytes
and 96.0% (48 out of 50) in vitro-matured oocytes.
The M-II karyoplast was removed successfully in 35 of 41(85.4%) of the donor oocytes and 40 of 48(83.3%)of the recipient oocytes. All of 35 karyoplasts of recipient oocytes were replaced in the periviteleine space of
enucleated donor oocytes and 28 of these 80.0% were fused to form a reconstituted oocyte. The fertilization
rate, cleavage rate and blastocyst formation rate following ICSI for constructed oocytes and recipients oocytes
were [77.1%(27/35), 65.7%(23/35), 25.7%(9/35)], [59.0%(58/98), 26.1%(25/98), 3.4%(3/98)] respectively.
Chromosomal analysis of 4 embryos following nuclear transfer indicated they were all diploid sets of 46
chromosome.
In conclusion, it has been demonstrated that oocytes constructed following the karyoplast transfer of in-vitro
matured M-II oocytes into enucleated freshly ovulated M-II oocytes clearly had more efficient andchromosomally normal embryonic development than did in-vitro matured oocytes after ICSI. These results
demonstrate that this technique can be applied to the treatment of female infertility due to ooplasmic
deficiency and abnormalities in aged oocytes.
◆Biosketch
Name: Atsushi Tanaka, M.D. Ph.D.
Date of Birth: December 22, 1949 - Niigata, Japan184

●Concurrent Symposium C-17.3

Education:
1970-1976 Juntendo University School of medicine, Tokyo Degree: M.D.
1976 Juntendo University Hospital, Department of Obstetric and Gynecology
1978-1982 Juntendo University Graduate School of Medicine
1982 Juntendo University Hospital, Department of Pathology
1983 Koshigaya city hospital, Chief Doctor of Obstetric and Gynecology
1990 Saint Mother Hospital, Director

185

●Concurrent Symposium C-18.1

TESE and Male Infertility
Sherman J. Silber
Infertility Center of St. Louis
Objective: To determine the relative impact, if any, of the degree of spermatogenic
defect, and sperm origin (testis, epididymis, or ejaculate), on pregnancy and delivery
rates with ICSI.
Design: A prospective study of unselected consecutive IVF cycles was conducted in a single center where allcycles were performed with ICSI regardless of the degree of male factor if any.
Materials & Methods: 719 consecutive cycles of ICSI with sperm retrieval were performed for azoospermicmen, whether obstructive (OA) or non-obstructive (NOA). During the same time interval, 1,849 consecutive
cycles of ICSI were performed using ejaculated sperm for couples with normal spermatogenesis as well aswith varying degrees of male factor defects. Conventional rather than minimal stimulation was used for allthese cycles.
Results: The age of the wife and the number of eggs retrieved had the greatest effect on pregnancy anddelivery rate for every sperm category evaluated, and sperm category had no effect on results. For ejaculated
sperm, whether severe oligospermia (less than 5x106), normospermia, or even donor sperm, pregnancy ratevaried only from 42% to 47%. However, pregnancy rates varied significantly from 54% to 18% related to theincreasing age of the wife, and to her decreasing ovarian reserve.
Testicular sperm (TESE) gave poorer results than epididymal (MESA) or ejaculated sperm, even when there
was normal spermatogenesis. The pregnancy rate for epididymal sperm (MESA) with obstructive azoospermia(OA) was 55%, but for TESE for OA was 38% (p<.001), and with TESE for NOA was 37%.
Conclusions: The degree of spermatogenic failure has no impact on pregnancy or delivery rate with ICSI.
However, proximal motile epididymal sperm and motile ejaculated sperm using ICSI gave consistently better
pregnancy and delivery rates than testicular sperm. There may be some aspect of sperm maturation which
occurs when sperm leave the testis that is beneficial for ICSI.
◆Biosketch
Dr. Sherman Silber is one of the world's leading authorities on IVF, microsurgery, vasectomy and tubal
reversal, egg and embryo freezing, ovary and testis transplantation, and the reproductive biological clock. He
is director of the Infertility Center of St. Louis at St. Luke's Hospital in St. Louis, and author of thebest-selling How To Get Pregnant.

186

●Concurrent Symposium C-18.2

Microdissection Testicular Sperm Extraction (MD-TESE)
Atsumi Yoshida, M.D., Ph.D, MBA
Kiba Park Clinic
Introduction
About 20% of men who visit an outpatient clinic or department of male infertility
have azoospermia. Of such patients, about 20% have obstructive azoospermia (OA),
while about 80% have non-obstructive azoospermia (NOA). NOA was considered to be absolute infertility inthe past; however, a case of pregnancy achieved by ICSI using testicular sperm from a man with NOA wasreported in 1995. Thus, even for men with NOA, pregnancy has become achievable if a trace amount ofsperm is present in the testes. In the past, a greater volume of testicular tissue than required was extracted toretrieve sperm by the conventional method (conventional testicular sperm extraction [C-TESE]). Recently,
however, microdissection testicular sperm extraction (MD-TESE) has been developed to retrieve sperm
efficiently (high sperm retrieval rate) and safely (low rate of adverse reactions), without extracting testicular
tissue at random. At this symposium, I would like to present the results of MD-TESE-ICSI that have been
performed at our clinic.
Results of MD-TESE
Azoospermia was diagnosed if no sperm is detected in the sediment in centrifuge tubes after centrifugation on
repeated semen tests. MD-TESE was performed in men with NOA, and the rates of retrieval of ―genuine‖
sperm (excluding pseudo-sperm and late-stage spermatids) according to the cause of NOA were as follows:
28.7% (109/380) of 46,XY men with NOA of unknown cause; 45.9% (34/74) of patients with Klinefelter‘s
syndrome; 50.0% (5/10) of patients with spinal cord injury; 77.8% (7/9) of patients with cryptorchidism; and45.5% (5/11) of patients after exposure to chemotherapy.
Klinefelter’s syndrome
Klinefelter‘s syndrome is the most common chromosome disorder among chromosome disorders associatedwith male infertility. At our clinic, ICSI or frozen-thawed embryo transfer was performed in 33 cases (77cycles), in which viable sperm could be detected by MD-TESE, and 32 babies (18 male and 14 female babies)
were born to 20 couples (25 cycles), and one woman is now in pregnancy. The fertility (live birth) rate forcouples with Klinefelter‘s syndrome in whom sperm was detected was 60.6% (20/33).
Cryptozoospermia
When semen tests are performed on separate occasions, no sperm is detected on one test (suggestive ofazoospermia), and several spermatozoa are detected in semen on another test. Such case is referred to as
cryptozoospermia, implying that sperm lies hidden (= crypto) in semen. In the case of cryptozoospermia, any
good embryo may not be obtained by ICSI using ejaculated sperm, or any good sperm may not be obtained
from ejaculated semen. In such cases, testicular sperm extraction may be performed in order to find better
sperm, or in an emergency on the day of egg collection. In addition, instead of conventional TESE performed
for OA, MD-TESE is usually performed to collect spermatozoa present in large, white seminiferous tubules,
and ICSI is then performed using the sperm thus collected. At our clinic, the sperm retrieval rate for men withcryptozoospermia was 83.3% (5/6).
Conclusion
MD-TESE-ICSI is an infertility treatment that relies heavily on the skills of physicians (reproductive
specialists: urologists, obstetricians, and gynecologists) and the skills of embryologists, that is, the total
strength of a fertility clinic.

187

●Concurrent Symposium C-18.2◆Biosketch
Mar. 1986
May 1986
Apr. 1992
Oct. 1993
Jul. 1994
Nov. 1997
Jan. 1999
Apr. 2007
Oct. 2009
Mar. 2011

Graduated from Ehime University School of Medicine
Department of Obstetrics and Gynecology, Tokyo Metropolitan Police Hospital
Japan Society of Obstetrics and Gynecology-certified specialist
Adjunct physician at Central Clinic (Tokyo)
Postgraduate researcher at the First Department of Urology, Toho University School of
Medicine
Received the degree of M.D. (Toho University)
President of Kiba Park Clinic
Certified as a supervisory doctor of reproductive medicine
Certified as a specialist of clinical genetics
Received the degree of Master of Business Administration (MBA) (Nihon University)

Kiba Park Clinic
Kamei Build 2nd Floor, Kiba, Koto-Ku, Tokyo, Japan, 135-0042

188

●Concurrent Symposium C-18.3

IMSI, already 8 years in ART practice. Where do we stand?
P. Vanderzwalmen1,2, G. Cassuto3, T. Neyer1, A. Stecher1,
M. Zintz1, N. Zech1
1
IVF Centers Prof. Zech - Bregenz, Austria
2
Centre Hospitalier Inter Régional Cavell (CHIREC),
Braine l‘Alleud, Belgium
3
Laboratoire Drouot, Paris, France
The implementation of Intracytoplasmic Morphologically Selected Sperm Injection (IMSI) was reported to be
associated with higher blastocyst, implantation and pregnancy rates and lower miscarriage rates (Bartoov,
Berkowitz, Antinori, Cassuto, Vanderzwalmen).
However, almost ten years after the introduction of IMSI, this approach of sperm selection still receives fierce
criticisms and scepticisms.
The selection of a normal sperm head morphology at high magnification prior to ICSI permits to select not
only normal spermatozoa based on shape and size criteria but in addition spermatozoa devoid of sperm headlarge vacuoles (LV)
Are there justified reasons not to select spermatozoa, especially such showing no LV?
1. Effectiveness of selecting a normal spermatozoon.
Selection of spermatozoa without vacuoles by IMSI was shown to be effective in case of:- repeated failure of implantation (Bartoov, Berkowitz, Antinori)- male infertile patient, (Bartoov, Berkowitz, Balaban)- high degree of DNA fragmentation (Hazout)- No blastocyst formation or low rates of blastocysts after ICSI (Vanderzwalmen, Cassuto).
2. Pathological character of nuclear vacuoles:
The formation and presence of large sperm head vacuoles is not a physiological process and thus needs
attention.
In most cases (60%), LV represent a nuclear thumbprint-like concavity associated with a complete or partial
failure of chromatin condensation predisposed to DNA damage secondary to a possible lack of DNA
methylation or abnormal histone acetylation and methylation (Garolla, Perdrix,Cassuto).
This concavity isischaracterized as a very thin area where the plasma membrane is intact but sunken(Boitrelle).
3. Their origin:
Abnormal sperm morphology is an indicator for an impaired progression of spermatids throughspermiogenesis (Franken). Meiosis abnormalities may affect the various steps in spermiogenesis that could
lead to insufficient chromatin condensation and perhaps immaturity of the sperm plasma membrane (Ovari,
Perdrix, Boitrelle, Cassuto) leading to the formation of LV (Miller).
4. Effects on embryo development and the health of the progenies
As nuclear defects such as LV negatively influence the outcome of embryo development due to theirimplicated late paternal genome effect, the pending question relates to their possible negative effects on thehealth of the progenies.

189

●Concurrent Symposium C-18.3

Recently Cassuto communicated the results of the first prospective study including 450 children born after
IMSI compared with the outcome of 578 children born after ICSI in a group of women younger than 39 years.

The number of major congenital malformations and genetic disorders, mainly affecting the genitourinary
system, was 24 from 578 ICSI (4.15%) versus 8 from 450 IMSI (1.77%) (X2 = 0.031) thus emphasizing theimpact of the sperm head morphology defects on congenital abnormalities.
5. How to appreciate the benefit of IMSI?
Additional to the former mentioned increase in implantation and pregnancy rates and probably be of benefit
for the health of offspring, IMSI was reported to increase the blastocyst rates. With the advanced vitrification
techniques nowadays surplus blastocysts are cryopreserved for later cycles and IMSI thereby contributes toincrease cumulative pregnancy rates (Vanderzwalmen)
6. Technical aspect
Some are reluctant to apply IMSI because as compared to ICSI it is more time consuming and more expensive.
Such arguments are not sustainable because our final aim is to help patients who give all their hopes.
◆Biosketch
Pierre Vanderzwalmen, embryologist, is currently working as scientific coordinator at the IVF center of Prof
Zech in Austria and in the CHIREC IVF institute of Prof Lejeune in Brussel.
His scientific investigations focused: on the relationship between selection of spermatozoa and embryo
selection criteria on the outcome of embryo development, and on vitrification of oocytes and embryos.

190

●Concurrent Symposium C-19.1

Clinical application of AMH in the reproductive medicine
Yoshimasa Asada1,2,3
1
Asada Ladies NAGOYA Clinic, Nagoya, Japan,
2
Asada Ladies KACHIGAWA Clinic, Aichi, Japan,
3
Asada Institute for Reproductive Medicine
Introduction: Japanese people are marrying later in life. Ovarian reserve is now
a important competent of evaluation for patients. Because of FSH levels insufficient to evaluate their ovarian
reserve, AMH has recently been seen as an alternative marker for such patients. Three years ago, I began
measuring the serum AMH levels in my patients and have reported on the importance of this hormone as amarker of ovarian reserve in reproductive medicine.
1) What is AMH? AMH is characteristically not modified through the menstrual cycle. Large amounts of
AMH are secreted from the granulosa cells of the preantral and antral follicles. Its secretions are decreased by
the maturing procedures of follicles. By such characters, the hormone is a quite useful marker for infertile
treatment. All patients visiting in my clinics are checked serum AMH level. The method for infertility
treatment is decided by this result. Although there are standard and mean AMH levels, there is no range seems
to be normal. Mean levels in women in their early 20s about 7 ng/ml, however its levels linearly go down 0 in
women in their late 40s. The ages of our infertile patients are not statistically correlated with AMH levels.
AMH levels, moreover, show irregular distributions not only in infertile patients but also in pregnant womenand fertile women.
2) AMH and ART: Previously in my Clinic, the ovarian reserve is evaluated by basal FSH, the number ofantral follicle, age, the patient history and outcomes of prior treatment. However, AMH levels are particularly
well correlated with the number of eggs can be collected. Convincingly, AMH is an excellent indicator of thenumber of collectable eggs in ART procedures. In my opinion, the AMH levels are most important to selectappropriate procedures of controlled ovarian stimulation for my patients. Although I also consider age, I havefound that patients with AMH levels of below 1 ng/ml, are candidates for simple ovarian stimulation because
they do not respond well to the short protocol. The patients with beyond 3 ng/ml of the AMH level seem torespond well to the antagonist protocol. A high percentage of quality ova are required for a high conception
rate. High AMH levels and a large number of ova increase the quality of eggs can be available in theoperations. While high AMH levels facilitate conception, pregnancy quality is largely dependent on age,meaning that AMH cannot be used to predict conception rates. Moreover, AMH is a very effective predictorof ovarian hyperstimulation syndrome and should be measured to prevent this side effect.
3) Current infertility treatments: I emphasize that mature egg can be collected in the operation bychecking AMH levels and the patient age, not by FSH. The conventional by-the-book decisions are no longer
effective because of this AMH method.
Conclusions: By measuring AMH levels in many patients, I realize that there is no mean range consistentenough to indicate ovarian age. Women cannot foresee their reproductive potential by their age. AMH is the
only true indicator. It can be concluded that the range of reproductive years are different in person. I canadvice women, whether married or unmarried, is to check her AMH level to know their true reproductive age
and obtain data to decide when she should be married and pregnant.

191

●Concurrent Symposium C-19.1◆Biosketch
Current position:
Executive Director, Asada Ladies'ClinicDirector,
Asada Ladies Nagoya Clinic Director,
Asada Ladies Kachigawa Clinic
Education and professional experience:
March 1982
Graduated from Nagoya University School of Medicine
May 1991
Worked as a staff member in the Department of Obstetrics and Gynecology, Nagoya
University School of Medicine
January 1993 Studied abroad at the Jones Institute for Reproductive Medicine, Eastern Virginia Medical
School, Norfolk, Virginia, where I performed basic research on ICSIMarch 1995
Began treating patients with ICSI at Nagoya University School of Medicine, Branch Hospital
May 1995
Published a paper reporting Japan's first pregnancy with ICSI using testicular spermatozoa
April 1998
Served as Director of the Infertility Center at Nakajima Clinic
April 2004
Founded Asada Ladies Clinic
April 2006
Received certification as Board Certified Member of Japan Society for Reproductive
Medicine
August 2010 Founded Asada Ladies NAGOYA Clinic

192

●Concurrent Symposium C-19.2

Chronological Aging vs Biological Aging: AMH as an early marker ofovarian aging
Budi Wiweko, Dyah MP Prawesti, Andon Hestiantoro,
Kanadi Sumapraja, Muharam Natadisastra
Division of Reproductive Endocrinology, Department Obstetrics and Gynecology,
Universitas Indonesia, Jakarta, Indonesia
BackgroundAs women aged, her ability to produce ovum with good quality and quantity will be decreased. This has been
related to chronological age with the ovarian biological age, representing the ovarian reserve and its response
to ovarian stimulation. Therefore this study was conducted to evaluate the correlation between thechronological age and ovarian biological age with a graph model and normogram of AFC, AMH, and FSH,
and to see the decreasing pattern of each variable based on age of the women.
MethodA retrospective cohort study with AFC, FSH, and AMH serum level data taken from medical records of IVF
patients at Yasmin Clinic, dr.Cipto Mangunkusumo Hospital, Jakarta, Indonesia between January 2008 to
December 2010.
ResultCorrelation between 3 variables, AFC, AMH, and FSH, related to the age is statistically significant. AFC andAMH serum level in graph with percentile 3, 10, 25, 50, 75, 90, and 97 has decreased following age, whereas
FSH increased following age. There is relatively lower sloping degree of FSH showed that it is increased inolder age compared to AFC and AMH, therefore FSH is observed to be a later predictor for evaluating ovarian
reserve, whilst AMH is an earlier predictor.
Conclusion
Age-related normograms in infertile women demonstrate a biphasic pattern of decreased antral follicles while
AMH and FSH transformed with a linear pattern. AMH found to be an earlier predictor for ovarian biologicalage assessment. These curve models and normograms could provide a reference guide for physicists tocounsel infertile women. However, future validation with longitudinal data is still needed.
◆Biosketch
Budi Wiweko is an OBGYN Consultant on Fertility currently working at dr. Cipto Mangunkusumo Hospital,
Jakarta, Indonesia and involved in Yasmin Reproductive Clinic dealing with patients with menstrual andfertility problem since 2006. He finished his study at Universitas Indonesia as an OBGYN in 2005 and had
been studying as a research student for 3 months at Hyogo College of Medicine, Nishinomiya, Japan back in2006. He continued his study as a consultant, and finished in 2008. He is interested in doing research focusing
on reproductive endocrinology and already wrote many papers.

193

●Concurrent Symposium C-19.3

Premature Ovarian Failure (POF) and Anti-Mullerian Hormone (AMH)
Bunpei Ishizuka, M.D.
Dept. Ob/Gyn, St. Marianna University School of Medicine
It has been thought that, once the diagnosis of POF is established, it is impossible toinduce ovulation except for rare, occasional spontaneous remission. We have been
attempted ovulation induction by giving estrogen- progestin replacement and rFSH/ HMG in POF patients.
AMH has often been reported as a predictor of ovarian reserve in infertility treatment. In the present study, we
examined the usefulness of AMH for predicting possibility of ovarian follicle growth and/or oocyte retrieval
in POF patients.
AMH levels were measured in 270 POF patients who started ovulation induction in 2010. Their AMH levels
ranged from 0 to 13.8 pmol/ml (mean 1.51±1.92 pmol/ml). We divided the patients into 4 groups; Group A,
patients in whom follicular growth was not observed by ultrasound or follicles did not reach the size that we
could attempt oocyte retrieval during the course of treatment; Group B, patients in whom follicular growth
was observed and oocytes were confirmed at oocyte retrieval but could not get fertilization; Group C, patientsin whom follicular growth was observed and oocytes were confirmed at oocyte retrieval and embryo was
obtained but could not get frozen embryo; Group D, patients in whom, frozen embryo was obtained. Group E;
patients in whom, no follicle growth was observed. We analyzed the AMH levels in these 5 groups of POF
patients to examine whether measuring AMH levels in POF patients could predict outcome of controlled
ovarian stimulation- IVF-ET.
◆Biosketch
1971
1979
1979-81
198219932000-present

M.D.: Showa University School of Medicine
Doctor of Medical Science: Keio University
Research Fellow: Dept. Reproductive Medicine, University of California, San Diego, U.S.A.
Assistant Professor, Dept. Ob/Gyn, St. Marianna University School of Medicine
Associate Professor, Dept. Ob/Gyn, St. Marianna University School of Medicine
Professor and Chair, Dept. Ob/Gyn, St. Marianna University School of Medicine

194

●Concurrent Symposium C-19.4

Dynamic serum AMH levels and AMH-related gene expression
in human ovarian follicles
Hsin-Fu Chen, Hong-Nerng Ho
National Taiwan University Hospital, Dept. of OB/GYN, Taipei, Taiwan
Introduction: The predictions of ovarian reserve and probability of pregnancy afterin IVF or ICSI are significantly relevant in clinical practice. An accurate prediction can efficiently improvecounseling for patients, selecting optimal ovarian stimulation protocols, preventing unfavorable outcomes(including ovarian hyperstimulation syndrome or cancellation of treatment), and predicting the pregnancy
outcome. Commonly used methods for predicting ovarian reserve include baseline FSH and inhibin B levels,
antral follicle count (AFC), and in recent days serum AMH levels. It was reported that although FSH, inhibin
B and AFC all could predict the ovarian response after gonadotropin stimulation, only AMH might predict the
pregnancy outcome. However this assumption remains to be tested and the AMH-related gene expression infollicular compartment has been less well explored. The development of mature eggs in the ovarian follicles is
a complex and ill-defined process which involves somatic cells in the follicles to nurture oogenesis. Recent
studies also revealed critical roles of the oocyte itself in folliculogenesis and showed the intimate bidirectional
communication between the oocyte and somatic cells. It has been presumed that some factors may participate
in this communication and these may include, though not restricted to, KL, AMH, Activins, GDF9, FSH,
BMP15 and LH. Previous study has shown that both thecal and granulosa cell dysregulation may occur inpatients with PCOS, which lead to the extreme ovarian response during ovarian stimulation. It showed that
AMH, AMH-receptor II, FSHR, and AR genes were over-expressed by granulosa cells from women with
PCOS. Recent report also showed that level of AMH in the follicular fluid, but not in the serum was a more
useful marker of subsequent embryo implantation.
Materials and Methods: We therefore search the literatures for this issue. In addition we examined the serial
serum AMH levels of patients undergoing IVF program and collected the follicular fluid for AMH and steroid
hormone levels. The luteinized granulosa cells from smaller and dominant follicles were also separatively
collected for the examination of the AMH, AMH receptor, AR, FSH receptor, and LH receptor transcriptlevels by real-time RT-PCR. This talk therefore aims to clarify the role of AMH-related gene expression in
relation to human reproduction by analyzing the above mentioned data.
Results: Our data showed that the serum levels of AMH showed a trend toward decrease at an insignificantly
manner during the course of ovarian stimulation in patients undergoing IVF. A nadir could be identified
around the time of hCG injection and started to return gradually to the baseline level at the time of pregnancy
checkup. Subsequently we will also show the data comparing patients with different ovarian responses to theovarian hyperstimulation.
Conclusions: Intrafollicular factors including AMH significantly modulate the ovarian responses during
ovarian hyperstimulation. It is likely that identification of the signals originating from these factors may
improve our clinical practice in ART.
◆Biosketch
Dr. Chen obtained his M.D. degree from National Taiwan University (NTU) and received resident training in
the Department of OB/GYN, NTU Hospital (NTUH) (1988-1992). He received fellowship training for
Reproductive Endocrinology and Infertility in NTUH (1992-1994), did postdoctoral research in the
Department of OB/GYN, University of British Columbia, Canada (1996-1997), and received short-term study
for infertility treatment in Assistance Hopitaux Publique De Paris, Cochin (Universite De Paris V). Dr. Chenmajors in the treatment of diseases related to reproductive endocrinology and infertility. His research interests
are focused on embryo growth and the development of pluripotent stem cells to germ cells. He is now theassociate professor/visiting staff in the Department of Ob/Gyn and Graduate Institute of Clinical Genomics,
NTUH. Dr. Chen is also the President of Taiwanese Society for Reproductive Medicine (TSRM).

195

●Concurrent Symposium C-20.1

Implementation of the fertility assistance program after the Wenchuan
earthquake in China
Shangwei Li, Xiaohong Li, Lang Qin, Han Hu, Shan Luo, Lei Li
West China Second Hospital, Sichuan University, Chengdu Sichuan, China
Objective: To report the implementation of the fertility assistance program after the Wenchuan earthquake in
Sichuan, China.
Methods: Fertility assistance program has been carrying out for these parents who lost children in the
Wenchuan earthquake since 8, 2008, 3 months after the quake in China. This program included 3 parts:sociomedical fertility health support; psychological counseling and intervention in reproductive health; and an
assisted reproductive technology (ART) service.
Results: There were over 8000 couples who lost their only child in the earthquake and planned to haveanother one, but their reproductive health and psychological well-being were not well in the following several
months. Our observation study found that about 30% (59/197) of the surviving women with a high degree ofloss in the earthquake reported menstrual irregularity within 6 months after the earthquake. Association
analyses showed that some stressors of the Wenchuan earthquake were strongly associated with self-reports ofmenstrual irregularity, including the loss of children (RR: 1.58; 95% CI: 1.09, 2.28), large amounts ofproperty (RR: 1.49; 95% CI: 1.03, 2.15), and social resources (RR: 1.34; 95% CI: 1.00, 1.80). Our study also
reported that 44.7% (88/197) women were in the status of anxiety and 91.9% (181/197) women had
depression within 6 months after the earthquake, which were significantly higher than those in the control
groups (P&lt0.05). With the help of the fertility assistance program, majority of the surviving women restored
good reproductive health and psychological well-being. There were 3,761 women achieved successful
pregnancies and 2,864 healthy babies were born up to 3, 2011. For those couples with fertility problems, wegave medical interventions such as medcines, surgeries, and ART. The major reproductive problems ofsurviving women older than 40 years old were low pregnancy rate and high spontaneous abortion rate, whichwere very challenging for our post-earthquake fertility assistance program.
Conclusion: The post-earthquake fertility assistance program played a great role in helping surviving womenreconstruct their lives and families.
◆Biosketch
ShangWei Li, professor of medical science and doctoral supervisor, is the chief of Assisted Reproductive
center of West China Second Hospital, Sichuan University, China. Her academic field involves assisted
reproduction technology, Family Planning and infertility. Her professional affiliations includes the member ofstanding committee and the vice chief of management and ethics group of the Chinese Society of
Reproductive Medicine, Chinese Medical Association; vice director of Sichuan Family Planning Policy
Association; academic and technical leader in Sichuan province, etc. She honored 38 red-banner pacesetter ofSichuan province and the national medical health system advanced individual in 2010. She rewarded 2 second
prize for technology progress of Sichuan province and the national patent for utility model. She undertakes
more than ten national and province based scientific research programs and has already tutored 20
post-graduate master students and 20 doctoral students.

196

●Concurrent Symposium C-20.2

Cross-border Cryo-shipping of Vitrified Embryos(CCVE): The newest
ART success story
Goral N Gandhi, MSc1, Gautam N Allahbadia, MD, DNB, FNAMS2,
Aaisha Khatoon, MSc3, Sakina Kagalwala, MSc4,
Gouri Gupta, MD, DNB5, Rubina Merchant, PhD6
1
Laboratory Director, Rotunda- Center for Human Reproduction, Mumbai,
India
2
Medical Director, Rotunda- Center for Human Reproduction, Mumbai, India
3
Embryologist, Rotunda- Center for Human Reproduction, Mumbai, India
4
Embryologist, Rotunda- Center for Human Reproduction, Mumbai, India
5
Consultant Fertility Specialist, Rotunda- Center for Human Reproduction, Mumbai, India
6
Embryologist, Rotunda- Center for Human Reproduction, Mumbai, India
Objective: To compare survival rates, implantation rates and pregnancy rates of locally vitrified humanembryos versus cross-border cryo-shipped vitrified embryos transferred into Indian gestational surrogates.
Design: Prospective clinical study
Materials and Methods: A total of 82 frozen-thaw embryo transfer cycles using gestational surrogacy
performed in our private clinic during a one year period from January 2010 to December 2010 were analyzed.
Group A included the frozen embryo transfers (FET) using locally vitrified embryos (n = 28) and Group B
included FETs using cross-border cryo-shipped vitrified embryos (n = 54). The KitazatoTM vitrification kit(Kitazato India, Mumbai, India) and CryotopTM device (Kitazato India, Mumbai, India) were used to
cryopreserve all the locally vitrified embryos. The cryo-shipped embryos were vitrified using different media
and devices.
The embryos were cryo-shipped to our center using the Rotunda CryoShipTM Global service, our own
cryo-shipping program, using a 21 day dry shipper. The shipper was exempt from X-ray screening at allinternational cross-border custom points. The shipper was tracked on the internet throughout its journey acrossthe globe, not just by the courier company but also by our embryologists to ensure successful, timely
deliveries. The infectious diseases screen of the commissioning parents at the time of embryo freezing & thegestational surrogate prior to Embryo Transfer were reviewed. All the embryo transfers were performed by
the same clinician in screened gestational surrogates aged 21-35 years.
Results: 94.51% of locally vitrified embryos survived as compared to 92.37% of cryo-shipped vitrified
embryos (p > 0.05). The locally vitrified embryos had 43.49% implantation rate and 53.85% clinicalpregnancy rate following transfer into gestational surrogates, as compared with 41.28% implantation rate and50.0% clinical pregnancy rate following transfer of cryo-shipped vitrified embryos into gestational surrogates(p > 0.05). There were no significant differences in all corresponding values. The Student's t-test,
Mann-Whitney test and Fisher's exact test were used where appropriate.
Conclusion: No significant difference was found between the viability of locally vitrified embryos and thecross-border cryo-shipped vitrified embryos. Hence we conclude that cross-border cryo-shipping of embryos,
when done appropriately, is a very efficient way to achieve a gestational surrogate pregnancy withoutcompromising the implantation and pregnancy rates.
◆Biosketch
Dr. Gautam Allahbadia

197

●Concurrent Symposium C-20.2

Gautam N Allahbadia is a noted world authority on ultrasound guided embryo transfers and one of the pioneers
in third party reproduction in South-East Asia. He has over 100 peer-reviewed publications to his credit and ison the Editorial Board of several International Journals. Dr Allahbadia was responsible for India's firsttrans-ethnic surrogate pregnancy involving a Chinese couple's baby delivered by an unrelated Indian surrogate
mother. He also has to his credit India's first Same-Sex Couple pregnancy and delivery of twins. Dr Allahbadia
has had quite a prolific career -- he has authored over 100 scientific papers, 60 book chapters and 13 textbooks,
the latest of which is a comprehensive text, entitled "Embryo Transfer" available from Jaypee Medical
Publishers. He was awarded the AOFOG-Young Gynecologist Award for the year 1998. He was recently madea visiting lecturer to the University of Tel Aviv- Sackler School of Medicine - the first Indian to be given such a
position.

198

●Concurrent Symposium C-20.3

Cross-border Reproductive Care: Exploitation or Opportunity in the
Global Quest for a Baby
Daniel S. Seidman MD
Department of Obstetrics and Gynecology of the Chaim Sheba Medical
Center, affiliated to the Sackler Faculty of Medicine, Tel Aviv University,
Tel Aviv, Israel
Cross-border Reproductive Care indicates, according to the ESHRE Task Force on Ethics and Law, themovements by patients looking for infertility treatment from one country or jurisdiction where treatment isunavailable for them to another country or jurisdiction where they can obtain the treatment they need. It wassuggested that the terms ‗reproductive‘ or ‗procreative tourism‘ should be avoided because of their negative
connotations and that the neutral term ‗cross-border reproductive care‘ should be used instead. Multiple
authors have recently pointed out in the medical literature that cross-border reproductive care represents anurgent and challenging issue to tackle from medical, legal, psychological and ethical perspectives.
The main reasons patients seek cross-border reproductive care include the pursuit of high quality care,absence of local availability, differences in laws and regulations, cost, social support concerns and ethnic
needs.
The most important motivators behind the recent rapid expansion in the demand for Cross-border
Reproductive Care include: 1. New information technology allows easy dissemination of information on
reproductive procedures in foreign countries; 2. International travel currently more comfortable andaffordable; 3. IVF clinics in certain countries are now actively seeking patients from foreign countries throughadvertisements in airline magazines or international satellite TV channels; 4. Fertility centers now offer
convenient all-inclusive packages that include not only the reproductive procedures, but also flight tickets,
escorted transfer, hotels, interpreters and local recreational tours; 5. The internet allows a very cost-effective
and efficient way for patients to compare the foreign clinic's services and unique merits in terms of cost,expertise in reproductive technologies and local policies; 6. More patients seeking treatment forbidden by law
in the couple's own country or is inaccessible to the couple because of their demographic or social
characteristics. As this relatively new phenomena seems unstoppable, growing concerns remain in regard to
the lack of international regulations and quality assurance.
◆Biosketch
Daniel S. Seidman, MD, MCSc
 Co-director, IVF Unit, Assuta Medcial Center, Tel-Aviv (one of the largest IVF Units in the world
with over 7,000 cycles per annum).
 Senior physician at the IVF unit of the Division of Obstetrics and Gynecology at the Sheba Medical
Center, Tel-Hashomer, Tel-Aviv.
 Associate Professor at the Sackler School of Medicine, Tel-Aviv University.
 President of the Israeli Society of Contraception and Sexual Health.
 Board member and member of the International Scientific Committee of the European Society ofContraception and Reproductive Health (ESC).
 Director of the Annual Course on Human Reproduction, School for Advanced Degrees of the Sackler
School of Medicine, Tel-Aviv University.
 Editor, Update Journal of the Israeli Obstetrics and Gynecology.
 Editorial board member, Israeli Journal of Medical Sciences (IMAJ) and Journal of Pregnancy.

199

●Concurrent Symposium C-20.3







Trained in Advanced Endoscopic Surgery for three years at the Stanford Endoscopy Center, under thesupervision of Prof. Camran Nezhat.
Completed a fellowship in Reproductive Surgery at the Stanford Medical Center, Stanford, California.
Two times winner of First Prize in the American Association of Gynecological Laparoscopists(AAGL) "Golden Hysteroscope Competition".
Published over 200 research papers and several books mainly in the field of human reproduction andadvanced endoscopic surgery.
Member of the Examination Committee of the Israeli Ob/Gyn Review Board.

200

●Concurrent Symposium C-20.4

Oocyte donors: prevalence of genetic, infectious diseases and others
Coll O; Guillen J, Garcia D, Colodron M; Rodriguez A; Vernaeve V
Clínica EUGIN, Barcelona, Spain
Oocyte donation (OD) is a well established treatment for ovarian failure andfor some genetic diseases. Pregnancy rates achieved by OD are above 50% percompleted cycle. Despite its effectiveness, this technique cannot be offered in all
countries due to legal constrains. In some other countries donor availability is limited for several reasons suchas donor non-anonymity, lack of donors or to very limiting requirements. Different laws, guidelines, protocols
apply in different countries in the world.
In the last decade, the Spanish donation model has become one of the reference models and certainly is themost effective in numbers. Spain achieves more than half of all oocyte donations performed in Europe.
Several reasons explain this phenomenon: Spain has always been the world leading country in solid organ
donation and has a long lasting tradition, its laws are well defined and are oriented to facilitate OD programsand donors may be compensated according to the European donation law.
Despite this favorable scenario, large numbers of donors have to be recruited in order to properly select themaccording to laws and guidelines. In Spain, donors should be 18 to 35 years of age, should not carry genetic
disorders or infectious diseases and be in good health.
Screening of inherited disorders such as known family history, an abnormal caryotype, cystic fibrosis carriers(etc.), infectious diseases such as HIV, HBV, HCV, sexually transmitted diseases, gynecological or
psychiatric disorders among others is performed.
There is no published information regarding the prevalence of such problems among a ―normal‖ young
population of women.
Data will presented on the prevalence of these diseases of abnormal screening tests in a normal population thatare evaluated in Clinica Eugin. Our center is one of the largest OD centers in the world with thousands ofdonation cycles completed per year.
These data are of interest for general in order to establish the prevalence of these disorders in an ―unselectedyoung female population‖, and to establish the performance of each single screening test. But the percentage
and the reasons for excluding donors is as well an information of great value for fertility centers. It will allow
them to plan in advance the real number of donors and costs, required to achieve a certain number of OD.
Furthermore, once a disease or a carrying state is identified in a donor, the workload to complete confirmation
tests, and to treat or refer patients to another institution or physician (general practitioner etc. ), may as wellbe established.
◆Biosketch
Name: Oriol Coll, MD, PhD
Education: Physctian. Obstetrics and Gynecology
Current work position: Clinica Eugin. FounderAdress: Entença 293 baixos, 08029 Barcelona. Spain
Academic and Societies positions achieved:
MD and PhD: University of Barcelona
Obstetrics and Gynecology: Hospital Clinic Barcelona
Executive Education: General Management. IESE Business School.
Fellowship in Reproductive immunology and Infectious Diseases. University of California at San Francisco

201

●Concurrent Symposium C-20.4(1988-1990)
Prior positions:
Director of Clinica Eugin 1999-2010
Former head of High Risk Obstetrics at the Hospital Clinic, University of Barcelona (2003-2010)
Professor at the Universty of Barcelona (1990-2010)
Areas of interest:
Assisted Reproduction
Perinatal Medicine
Infectious diseases
Indexed scientific publications: 80

202

●Concurrent Symposium C-20.5

Vouchers for health: A demand side output-based aid approach to
reproductive health services. A model to pay for infertility services?
Claus P. JanischHealth Policy Department, IGES Institut
Reaching the United Nations Millennium Development Goals has been a focus formany countries and development partners. In many developing countries with low levels of development,
access to and equity of basic quality health services is limited, especially for the very poor. Among poor
populations, maternal mortality is high as access to medical care and financial means are lacking. Infertility
represents a national health problem in many African- and also poorer Asian countries. Limited financial
health resources in developing countries are a major obstacle facing infertility management. Affordable IVF
treatment for financially needy couples is, nevertheless, supported by many medical experts, human rightgroups, and religious leaders A new approach, herein referred to as Vouchers for Health, is proposed as apossible way to organize and finance infertility services for the poor. The way that these programs are being
implemented in a number of countries for women's health services demonstrates, that the voucher based
approach comprises a variety of key structural elements of a national health insurance scheme: accreditation;
quality assurance; reimbursement system; claims processing; integrating the public and the private sector;
client choice; provider competition; and access to and equity of services provided. Voucher Schemes ma be a
way to finance IVF services.
◆Biosketch
Born 1948 in Celle in the north of Germany Claus Janisch studied medicine from 1969 till 1975 at the free
University of Berlin in West Germany. He specialized in the field of Obstetric and Gynecology at the
Academic hospital in Berlin Neukölln (Professor Erich Saling), at the University Hospital Vienna (Professor
Eduard Gitsch), and at the University Hospital Mainz (Professor Friedmann). He then worked for a couple ofyears as senior consultant (Oberarzt at various major hospitals. In 1984 Dr. Janisch joint the 'Deutsche
Gesellschaft für Technische Zusammenarbeit' an organization of the German Government to implement the
support of the German Government with developing countries. Till 189 he was Director of three Ministry of
Health Hospitals in Tabuk, Hail and Najeran in Saudi Arabia. In 1989 Claus Janisch became the first Medical
Advisor of the German Development Bank (KfW Banking Group). He was responsible for the financing ofhealth projects in Asia, Africa and South America.

203

●Concurrent Symposium C-20.6

Global Challenges and Perspectives
Sheryl Vanderpoel
World Health Organization, Headquarters, Geneva, Switzerland
Introduction: No single country in the European Union and, few transitional anddeveloped Asian countries had a total fertility rate above replacement level in 2009.
Many are coming close to becoming, or already defined as, low fertility countries. Women in developed countries
are having fewer children, but this trend is expanding into countries in transition and urban centres in developing
countries. In some countries, with burgeoning youth populations, this can be viewed as a positive trend. However, ahealthy balance is required, as prolonged delays in childbearing can skew population compositions if a large
proportion of women begin having their children significantly later in life. Although initial population decline canprovide an economic boost, the ultimate long term sustainability of political, social and economic national models
can become constrained if low fertility population trends continue to stagnate and do not reverse. The generalterm, "infertility" is defined as a disease of the reproductive system which results in an inability to become pregnant
or bring a pregnancy to term. Primary infertility (no prior pregnancies) is predominantly a result of delayedchildbearing, complications of childbearing and/or congenital disorders. Secondary infertility (infertility
following a previous pregnancy or live birth) is often a result of maternal sepsis (infection following delivery) or ofcomplications associated with R/STIs (including HIV/AIDS) and unsafe abortions.
Results: The joint WHO/World Bank World Report on Disability (June 2011): Infertility following maternal sepsis
and unsafe abortion was ranked 8th highest in prevalence as a disability across all age groups. Among populations
under the age of 60, infertility ranked 5th highest in prevalence as a major global disability. Within public healthdocuments and strategies, "family planning" is often interpreted as "contraception" and emphasizes the preventionof unwanted pregnancies. Within other public health strategies, critical "maternal and child health" care issues
are emphasized and addressed - only when a woman presents, often far into, her pregnancy. Peri-conception care,which includes infertility care, is the critical link between these two areas of reproductive and maternal health careservices. Attempting to rectify sub-standard or poor "beginnings" which may occur at the time of conception by
providing later enhanced antenatal care may be "too much, too late." Peri-conception services together withevidence-based reproductive medicine clinical management and interventions can better ensure safe pregnancy andbirth and decrease use of socio-behavioural methods to solve infertility. Traditional methods, in the absence ofquality infertility care, are problematic on a global level within high HIV endemic areas; they exacerbate not only
the problem of horizontal (and vertical) transmission of infectious diseases such as HIV/AIDS and syphilis, but can
result in a further increase in infertility within the community.
Conclusion: Access to simple, available and affordable solutions for peri-conception and infertility care services
represents a highly neglected component of global public health care. Revitalization of the field of infertility andperi-conception care is critical in order to fully and adequately address the Secretary General's Global Strategy for
Women and Children's Health to ensure healthy mothers, healthy children and healthy family life.
◆Biosketch
Dr. Sheryl Vanderpoel,
Current: Scientist and Medical Officer in the Director's Office of the Department of Reproductive Health andResearch, World Health Organization, WHO-RHR; UNDP/UNFPA/WHO/World Bank Special Programme of
Research, Development and Training in Human Reproduction (HRP). Primary responsibilities: scientific, technical,
financial and ethical peer review of research proposals submitted to HRP; RHR/HRP's research and normative
work in peri- conception concentrating on issues associated with infertility; and activities in ethics in sexual andreproductive health and assisted reproduction; and representing WHO-HQ at inter-agency/international
consultations in these areas of focus.

204

●Concurrent Symposium C-20.7

Affordable IVF – Low cost IVF: The Arusha project
Willem Ombelet
Department of Obstetrics and Gynecology, Genk Institute for Fertility
Technology, Schiepse Bos 6, 3600 Genk, Belgium (coordinator of the
ESHRE Special Task Force on Developing Countries and Infertility)
INTRODUCTION: The majority of childless couples are residents of developing
countries. When compared to Western societies, negative consequences of childlessness are experienced to agreater degree. Residents of developing countries encounter a lack of facilities at all levels of health care but
especially infertility diagnosis and treatment. Tubal infertility due to sexually transmitted diseases, unsafe
abortion and postpartum pelvic infections are the main causes of infertility in developing countries. Thismeans that most cases of infertility are only treatable with assisted reproductive technologies (ART) which areeither unavailable or very costly.
METHODOLOGY: December 2007 an expert meeting was organized by ESHRE in Arusha, Tanzania. The
meeting was the start of a global project aiming at increasing the diagnostic and therapeutic options forchildless couples in developing countries, emphasizing the need for reproductive health care education. The
final objective is the implementation of infertility services in many developing countries if possible linked toexisting family planning and mother care services.
RESULTS: Pilot studies will be organized to study the results of the combination of a one-day diagnostic
phase, the use of clomiphene citrate for ovarian preparation and the use of an unexpensive high quality IVF
laboratory phase.
CONCLUSION: Although prevention is better than cure, we believe it is justified and possible to implement
simplified, safe and effective methods of ART in resource-poor countries. We propose a special designed
infertility care program leading to a cost effective simplified ART program as a valid treatment protocol indeveloping countries when prevention has failed.
◆Biosketch
Willem Ombelet started his career researching infertility and IVF in 1984 in Pretoria, South Africa. Since
1987, he has been working in the Department of Obstetrics and Gynaecology at the St. Jans Hospital in Genk,
Belgium. In 1998, he obtained his PhD degree at the University of Leuven on ‗The value of spermmorphology and other semen parameters in diagnosis and treatment of human subfertility. He became theHead of the Department of Obstetrics and Gynaecology at the St. Jans Hospital in Genk, in 1999. From 2001
until 2004 he was the President of the Flemish Society of Obstetrics and Gynaecology.
Willem Ombelet (co-)authored 89 international peer-reviewed articles and was a (co-)-editor of 18 books. He
received 2 international awards.
Dr Ombelet is also the founder of the Genk Institute for Fertility Technology.
He organised nine ―Andrology in the Nineties‖ meetings and different workshops on assisted reproduction.
He is a consultant and Professor (Reproductive Medicine) at the University of Hasselt and the University of
Ghent.
Since 2006 he is the coordinator of the ESHRE Special Task Force on ―Developing countries and infertility‖.
Presently he is the chairman of the Workgroup ―Reproductive Medicine‖ of the Flemish Society of Obstetrics
and Gynaecology since 2007 and a board member of the Belgian Society of Obstetrics and Gynaecology(BSRM) and the Belgian College of Reproductive Medicine.
Willem Ombelet is also the editor-in-Chief of the international scientific journal ―Facts, Views & Vision in
ObGyn‖.

205

●Concurrent Symposium C-21.1

How to improve implantation in ART: novel approaches.
Yael Gonen, MD
Infertility Clinic, Lin Medical Center, Haifa, Israel
Recurrent implantation failure is a distressing phenomenon, both for the infertile
couple and for physician responsible for their treatment. Aetiology is often not
clear and treatment options are vague. Particularly when transferred embryos are ofgood quality, recurrent implantation failure may be attributed to decreased endometrial receptivity, molecularabnormalities at the endometrial level and abnormal embryo – endometrium dialogue. Furthermore, there may
be over or under expressed genes that may be related to successful implantation or less than optimal embryotransfer technique and factors with combined effect.
At present, the diagnosis of recurrent implantation failure is not sufficiently specific and we do not have thetools to diagnose in each case the exact cause for the repeated failure. The expense, time and stress has led to asearch for new technologies and drugs that will increase success rates, however progress has been limited.
Novel approaches to treat this frustrating and limiting step in the success of ART will be presented.
◆Biosketch
Dr. Yael Gonen received her MD degree from the Sackler School of Medicine at the Tel Aviv University in
Israel and completed residency in OB/GYN at Lady Davis Medical Center in Haifa.
After two years of a clinical and research fellowship in the field of Infertility and Reproductive Endocrinology
with Prof. Robert Casper at Toronto General Hospital, University of Toronto in Canada (1987–1989) she
returned to Haifa to the OB/GYN department at Lady Davis Medical Center and performed as Senior
Physician in IVF Unit.
In 1996 Dr. Gonen founded the new IVF Unit at Herzliya Medical Center in Haifa, while maintaining theposition of Medical Director for over three years.
Presently Dr. Gonen holds the position as Head of Infertility Clinic at Lin Medical Center in Haifa in addition
to maintaining a private practice.
Her abundant experience, dedication and avid participation in the field of infertility has earned her awards,
recognition for both clinical and research accomplishments. Her scientific studies are quoted in leading
journals and international meetings.
Dr. Gonen pioneered the use of ultrasound for endometrial quality (pattern and thickness) related to ART
pregnancy, she was the first to publish in 1990, the use of GnRH agonist in place of HCG as a means totrigger the gonadotrophic surge and also pioneered the use of Oral Contraceptives for follicle synchronizationand cycle scheduling in IVF.

206

●Concurrent Symposium C-21.2

Studies on the efficacy of immunotherapy using paternal mononuclear
cells for patients with infertility
Koichi Takakuwa
General Center for Perinatal, Maternal and Neonatal Medicine,
Niigata University Medical and Dental Hospital, Niigata, Japan
Although the etiology of recurrent spontaneous abortion (RSA), defined as three or more consecutive earlypregnancy losses, is often unclear, several investigators have reported the occurrence of immunologicallyexplainable RSA. Immunotherapy for patients with RSA using the husband's mononuclear cells has beenreported for the past three decades. Although the efficacy of this modality has been controversial, therandomized trial revealed the efficacy for patients with RSA, especially for those who were negative forblocking antibodies evaluated by a mixed lymphocyte culture reaction between spouses (MLR-BAbs). In thispresentation, I show the results of immunotherapy for unexplained primary RSA using freshly preparedmononuclear cells from the husband and the efficacy of the therapy, especially in patients negative for
MLR-BAbs. Moreover, there is possibility that the immunotherapy could be effective for patients withrecurrent IVF-ET failure, as the recurrent IVF-ET failure is considered to be the similar condition as therecurrent early miscarriages. We previously tried to evaluate the efficacy of the immunotherapy for patientswith recurrent IVF-ET failure. So, in this presentation, the results of immunotherapy for recurrent IVF-ET
failure are shown, in addition to the results of immunotherapy for RSA.
Two hundred and twenty-eight unexplained RSA were prospectively followed up by means of
non-randomized study regarding the immunotherapy. Of 228 patients, 165 cases underwent theimmunotherapy (group I). No MLR-BAbs were observed in these patients prior to vaccination. Pregnancy
outcome was also analyzed in RSA who revealed positive MLR-BAbs without immunotherapy (group II).
Furthermore, the outcome of 20 pregnancies in 18 unexplained RSA was analyzed who were negative for
MLR-BAbs and had become pregnant without immunotherapy (group III). In group I, of 140 newly pregnant
patients after vaccinations, pregnancy successfully continued in 110; thus, the success rate of the therapy was
78.6%. Regarding group II, in 24 of 32 patients (75.0%), the pregnancy continued successfully. The rate ofcontinuation of pregnancy was not significantly different between groups I and II. Pregnancy was successfully
continued in 6 cases (30.0%) in group III. The rate of successful pregnancy in experimental groups I and II
was significantly higher compared with that in group III.
Fourteen patients underwent the immunotherapy concerning the immunotherapy for recurrent IVF-ET failure.
The selection criteria for the immunotherapy were as follows; 1) the patients experienced 3 or more IVF-ET
failures in spite of transfer of good quality embryos, 2) they were negative for autoimmune examinations suchas anti-phospholipid antibodies, 3) they were negative for MLR-BAbs. Of 14 immunotherapy patients, ET
was performed at 30 cycles, and successful pregnancy was achieved in 6 cycles (20.0%). As control, 20 caseswith same condition as immunotherapy group, and did not undergo immunotherapy, were analyzed. Of 20
cases, ET was performed at 48 cycles and pregnancy was achieved in 3 cycles (6.3%). The tendency of
improvement of pregnancy rate was observed in immunization group (20.0% vs. 6.3%, p=0.07).
In conclusion, the immunotherapy using the husband's mononuclear cells on RSA negative for MLR-BAbs iseffective, and there is possibility that the immunotherapy might be effective for patients with recurrent
IVF-ET failure.
◆Biosketch
2009-present: Professor and Director, General Center for Perinatal, Maternal and Neonatal Medicine, Niigata
University Medical and Dental Hospital207

●Concurrent Symposium C-21.2

1998-2009:
1994-1998:
1986-1994:
1979:

Associate Professor, Department of Obstetrics and Gynecology,Niigata University School of
Medicine
Assistant Professor, Department of Obstetrics and Gynecology,Niigata University School of
Medicine
Instructor, Department of Obstetrics and Gynecology, Niigata University School of Medicine
Passed the Examination of National Board of Medical Doctor

Societies:
Executive Board Member, Japan Society for Immunology of Reprodcution
Councilor, Japan Society of Obstetrics and Gynecology
Councilor, Japan Society of Perinatal and Neonatal Medicine
Councilor, Japan Society of Reproductive Medicine
Councilor, Japan Society for the Study of Hypertension in Pregnancy
Councilor, Japanese Society for the Sexually Transmitted Disease208

●Concurrent Symposium C-21.3

Improvement of implantation rates using autologous peripheral bloodmononuclear cells
Hiroshi Fujiwara, Yukiyasu Sato, Akihito Horie, Ko Suginami,
Hisanori Matsumoto, Ikuo Konishi
Department of Gynecology and Obstetrics, Graduate School of Medicine,
Kyoto University
Introduction: In humans, the implanting embryo secretes human chorionic gonadotropin (HCG) andstimulates the maternal corpus luteum of pregnancy via blood circulation to produce progesterone.
Progesterone in turn maintains embryo implantation in the uterus during early pregnancy. Along with thisembryo-maternal crosstalk by endocrine system, accumulating evidence suggests that circulating immune
cells also play an important role in embryo implantation.
Material & Method: The effects of human peripheral blood mononuclear cells (PBMC) and murine spleencells on embryo implantation were accessed by human luteal cell culture, human trophoblast invasion assay
and murine implantation experiments. The effects of human PBMC were also evaluated by clinical trials.
Results: PBMC derived from pregnant women increased the progesterone production by human luteal cells
and promoted the invasion of embryos in vitro. In mice, spleen cells derived from early pregnant mice could
promote endometrial differentiation and embryo implantation in blastocyst-transferred pseudopregnant micein vivo, suggesting that the maternal immune system undergoes functional changes by recognizing developing
embryos from the very early stages of pregnancy. In addition, recombinant-HCG increased chemokine
production by human PBMC through lectin-glycan interaction, promoting trophoblast invasion. Furthermore,
intrauterine administration of autologous PBMC that were treated with HCG effectively improved pregnancy
and implantation rates in patients with repeated implantation failure during in vitro fertilization therapy.
Conclusion: These findings suggest that circulating immune cells are useful materials to improve
implantation rates in ART.
◆Biosketch
I. Academic and Professional Backgrounds:
1983 Graduated from Kyoto University, Faculty of Medicine 1985 Medical Staff, Dept of OB/GYN,
Hikone City Hospital
1986 Director, Dept of OB/GYN, Otowa Hospital
1988 Medical Staff, Dept of OB/GYN, Wakayama Red Cross Hospital
1993 Assistant Prof, Dept of GYN/OB, Faculty of Medicine, Kyoto University
1995 Lecturer, Dept of GYN/OB, Faculty of Medicine, Kyoto University
2009- Associate Prof, Dept of GYN/OB, Faculty of Medicine, Kyoto University
II. Academic Degree Medical Doctor:
1983 Kyoto University Doctor of Medical Science
1994 Kyoto University
III. Academic Awards: Research Promoting Award of Japan Endocrinology Society (1997)
IV. Editorial Services
1. Human Reproduction (Associate Editor, 2001-2003)
2. Reproductive Biology and Medicine (Executive Editor, 2003- )3. Journal of Obstetrics and Gynaecology Research (Associate Editor, 2008-)
4. American Journal of Reproductive Immunology (Editorial Board, 2010-)

209

●Concurrent Symposium C-21.4

Prevention of infertility in patients for the antiphospholipid syndrome anddeficiencies of natural anticoagulants
Moritoshi Seki
Sekiel Ladies Clinic
Antiphospholipid syndrome is a multisystem disease with the major elements of venous and arterial
thrombosis, recurrent pregnancy loss, fetal death and infertility. New mechanisms are described by which
antiphospholipid antibodies could cause placental thrombosis and infarction, acting directly on the surface
anticoagulant expressed on trophoblastic cells. The treatments have been proposed to treat with low doses ofaspirin and preventive or effective dose of heparin.
Oocyte retrievals of 1992 cases (736 patients) and embryo transfer of cryopreservation of 841cases (460
patients) from 2008 to 2009 years in Sekiel Ladies Clinic were analyzed for the estimation of effectivenessheparin and aspirin in recurrent pregnancy loss.
Out of the 736 patients examined, 6 patients resulted positive for abeta2GPI, 2 patients for lupus anticoagulant
antibodies, 43 patients for antiphosphatidylserine IgG, 131 patients for antiphosphatidylethanolamine and 31
patients positive for anti-phosphatidylinositol antibody.
One patient for deficiencies of natural anticoagulants protein C, 2 patients for protein S and125 patients forthe decrease in factor XII activity.
There is increasing evidence that heparin exerts its effect by inhibiting complement activation rather than by
its anticoagulation capacity. We aim for improve the outcome of pregnancies in APS by the invention oftherapies using combinations of aspirin, unfractionated heparin and/or low molecular weight heparin.
We infuse unfractionated heparin with Coopdech Syrinjector (Daikenika CO., LT. Japan) that is disposable
infusion pump operated by the use of atmospheric pressure. Patients can take the infusion of unfractionatedheparin dealing 24 hours at home.
We analyzed 97 cases (35 patients) of recurrent miscarriages patients in 1992 cases (736 patients) of IVF-ET.
Embryo transfer was performed in 73 cases. Forty nine cases (29 patients) were treated by using combinationsof aspirin and unfractionated heparin (UFH). Eighteen cases were pregnant and 4 patients delivered. Twenty
five cases (11 patients) were treated by using combinations of aspirin and low molecular weight heparin.
Thirteen cases were pregnant and 5 patients delivered.
We analyzed 66 cases (24 patients) of recurrent miscarriages patients in 841cases (460 patients) of embryotransfer of cryopreservation. Thirty four cases (15 patients) were treated by using combinations of aspirin andunfractionated heparin, 17 cases were pregnant and 5 patients delivered 6 babies. Twenty five cases (11patients) were treated by using combinations of aspirin and low molecular weight heparin, 18 cases werepregnant and 7 patients delivered.
The chromosomal abnormalities were 88.9% in products of conception (POC) chromosome analysis. The
average age of patients of recurrent miscarriage is 38.6 years old. The early treatment is recommended toprevent the chromosomal abnormalities.
We stop the heparin treatment by 8-10 weeks gestation for the introduction of patients to birthing hospital.
Some of the patients should have been cared by keeping heparin even after 10 weeks gestation by following
up the subsequent history of the patients.
The women with recurrent pregnancy loss should receive the prophylactic-dose heparin and aspirin.
◆Biosketch
1974
graduate from the medical department of Toho University
1974-1976 Intern of Toho University
1978
Assistant of Department of Obstetrics & Gynecology in Akita University
1979-1981 Scholarship of Humboldt foundation, Department of Obstetrics & Gynecology in Kiel University
1992
Assistant professor Department of Obstetrics & Gynecology in Gunma University
2001
The director of Sekiel Ladies Clinic

210

●Concurrent Symposium C-22.1

PGD for recurrent pregnancy loss
Atsushi Tanaka
Saint Mother Hospital, Fukuoka, Japan
It has been commonly assumed that preimplantation genetic diagnosis (PGD) for
translocations in couples with recurrent miscarriages has little advantage overnon-PGD intervention. We did PGD for translocation in couples with recurrent miscarriages and compared theresults to natural conception. One blastomere from 8 cell embryos following IVF in 55 cycles (33 patients)
was biopsied by the extrusion method and normal type (normal chromosomal or balanced translocation) was
distinguished from the abnormal type (unbalanced translocation). A single normal-typed embryo wastransformed. Fifteen couples suffering translocation with recurrent miscarriages and not using PGD were
followed up for 2 years. Of 64 cycles of PGD (36 patients), 22 subsequently conceived (pregnancy rate:
37.5% [24/64]) per cycle, 66.7% [24/36] per patient), resulted in 18 live births with a miscarriage rate of
20.8% (5/24). Of 24 pregnancies, 15 conceived at the first PGD (62.5% [15/24]). Seven conceived at thesecond trial (29.2% [7/24]) and 2 at the third trial (8.3% [2/24]). All 24 cases achieved conception by the 3rdtrials. The average maternal age and the collected eggs number from 32 unsuccessful cycles which had been
performed more than three times were 39.2 yrs and 3.4 eggs. These figures were significantly lower than thatof 24 successful cases with an average maternal age of 33.2 yrs and an average collected egg sum of 9.8 eggs.
Of 15 cases, 10 were conceived without PGD, resulting in 6 live births (birth rate: 40.0%) with 3 miscarriages(miscarriage rate: 30.0%). The six cases had an average 2.7 miscarriages and took on average 12.3 months
until the live birth. In conclusion PGD is clearly advantageous for couples with recurrent miscarriages
suffering translocation when they are younger than 35 years of age and the collected oocytes are expected to
be over 10 in total. The next concern is whether it is possible to distinct between a balanced translocation anda normal karyotype in order to select the normal one correctly. After providing informed consent,
morphologically excellent 3-day-old human embryos (8-cell stage after IVF), along with immature GV
oocytes, were donated by our clinic's patients for this study. The immature GV oocytes were cultured
overnight until the first polarbody was extruded. The 3-day-old embryo and IVM oocytes were freed fromthe zona-pellucida with a 0.3% tripsin solution. The embryos were then placed in PBS (without Ca and Mgions) to separate the blastomeres. Then electrically stimulated (AC 15V, 2 sec and DC 1000V/cm, 50 μsec) in
295mM D-mannitol containing PVP, Ca2+ and Mg2+. Each hybrid cell was treated with hypotonic solution(30% fetal bovine serum) for 20 min, and then affixed to a chromosome slide using the gradual fixation-air
drying method. 22 human blastomeres and 22 IVM oocytes successfully combined through electricalstimulation to create 20 hybrid cells. Of these 20 hybrid cells 3 were unsuccessful, 13 contained 1 complete
set of clearly arranged meiotic chromosomes (n=23) that were derived from the oocyte. In conclusion thisassay system has the advantage that it only takes 2 hours from biopsy to chromosome analysis. In this study,
the success rate for chromosome analysis was as high as 65.0% (13/20) when the hybrid cells were preparedfor chromosome slides one hour after electro-fusion.
◆Biosketch(For Dr. Tanaka‘s Biosketch, please refer to his abstract for Concurrent Symposium C-17.3.)

211

●Concurrent Symposium C-22.3

PGD outcome and alternatives for single gene disorder
Kou Sueoka, Yasunori Sueoka
Keio University School of Medicine, Tokyo, Japan
Preimplantation genetic diagnosis (PGD) for single gene disorders is the principle oforiginal PGD aimed at preventing outbreak of a child for the client who is a genetic
carrier of the diseases. However, as for the PGD, always tailor-maid diagnostic chemicals and procedures are
required for various genotypes to be present in each individual disorder family. However, the methods ofgenetic diagnosis are going to the direction of common and stable analysis from tailor-maid diagnostic method
for individual genotype. On the aspect of the material collection for diagnosis, biopsy method is also changing
to be less invasive and according to the methods of genetic analysis. Especially blastocyst biopsy has been
performed for more accurate and with a higher potential diagnosis by obtaining many cells, which can afford
to apply for wide variety of genetic diseases, and also to collect enough DNA specimen of embryonic cells forgenome wide analysis. The amplification efficiency by WGA from single cell is not yet stable and enough foranalysis to the present. As for our PGD cases to date, there were structural chromosome aberration, deletion
type mutation, point mutation, triplet repeat diseases in Mendelian disorders as single gene disorders, andmitochondria disease as non-Mendelian disorders. The gene amplification is inevitable for a diagnosis ofsingle gene from single cell in the technical aspect. Nested PCR has been the basic procedure to amplify
single copy DNA up to enough level for analysis. However, it yield to amplify only a few target genes andunable to avoid amplification failure or DNA contamination. Whole genome amplification (WGA) has beendeveloped for genome wide analysis. WGA has several options with both gene amplification types of PCRbase and non-PCR base methods, which would be selected for the analytical method of amplified products.
WGA yields the difficulty of conditional setting of nested PCR, and enables a repetition diagnosis for an
amplification product. As a result to the present, WGA expands diagnostic width of gene genotypes andimproves to decrease dismissed embryos that the diagnosis is unable to finalize, whereas amplification
efficiency from single cell has not been yet more than that of nested PCR. In addition, WGA makes possible
to analyze both genes of compound heterozygote mutation stably. We are approaching PGD using WGA
method to the disorder gene carriers of compound heterozygote mutation gene such as Fukuyama congenital
muscular dystrophy (FCMD), and epidermolysis bullosa-muscular dystrophy syndrome, and also to myotonic
dystrophy (DM1) which is not easy to diagnose by PCR amplification on the expansion of CTG repeat length.
It is also a large benefit that array technology is available to apply for the diagnosis of this amplification
product. There is a new direction of PGS shifting to a diagnosis using comparative genomic hybridization(CGH) instead of uncertain florescence in situ hybridization (FISH). However, applying CGH or the otherarray analysis for PGD can give genome wide information from embryo, which might be including sporadic
mutations or chromosomal aberrations. Although genome wide technologies must be strong tools, the analysis
is not always only to the required genetic information. For this directionality, we need to consider and argue
on the aspect of genetic information handling in PGD.
◆Biosketch(For Dr. Sueoka‘s biography, please refer to his abstract for the Pre-Congress Workshop: PGD 1.)

212

●Concurrent Symposium C-22.4

Preimplantation genetic diagnosis for aneuploidy using wholegenome amplification and microarray comparative genomic
hybridisation
Alan Handyside
London Bridge Fertility, Gynaecology and Genetics Centre
The development of methods for whole genome amplification from single cells andmicroarray comparative genomic hybridisation (array CGH) have revolutionised our ability to detect copy
number abnormalities for all 23 pairs of chromosomes. Accurate analysis is now possible with polar bodies,blastomeres and trophectoderm biopsied from cleavage and blastocyst stages. Analysis of both the first andsecond polar bodies and corresponding zygotes has revealed multiple errors caused by premature predivisionof chromatids in women of advanced maternal age. This is further evidence that as women reach themenopause but before their ovarian reserve is exhausted, many and eventually all oocytes have multiple
aneusomies so that the resulting embryos are highly unlikely to be viable. Recent advances in the technology
will be reviewed which allow accurate detection of structural chromosome imbalance, particularly in carriersof balanced reciprocal and Robertsonian translocations.
◆Biosketch
Background in mouse and human embryology and genetics with over 200 publications. Created one of thefirst mouse transgenic mice using embryonic stem cells. With Prof Lord Robert Winston, achieved the firstpregnancies following IVF and PGD worldwide in 1990. First Chairman ESHRE Special Interest Group in
Reproductive Genetics and founder and first Chairman of the ESHRE PGD Consortium.

213

●Concurrent Symposium C-23.1

Human ES cell and iPS cell derivation: Clinical applications andbiological characterization
Hidenori Akutsu
Chief, Division of Stem Cell and Reproduction, Department of
Reproductive Biology,
National Research Institute for Child Health and Development
The potential to generate and expand human pluripotent stem cells in vitro has great implications for basic
research, drug discovery, toxicology, to study diseases and can potentially serve as a resource for cell
replacement therapies. The U.S. Food and Drug Administration (FDA) has recently approved the firstembryonic stem cell-based clinical trial, indicating that human application of stem cell research is being
realized. Regenerative medicine and cell therapy are based on the premise that large numbers of normal cells
and that the cells will migrate, integrate and survive for a sufficient length of time as to be clinically useful.
An important issue facing investigators irrespective of their cell of choice or target for therapy is growing cells
in sufficient numbers for treating patients in an aseptic environment with adequate safeguards, sterility andtraceability. As research translates to clinical therapies, one of the primary concerns is that therapeutic cells
may carry foreign pathogens or antigens that induce disease or immune response by the patient. These
undesirable byproducts may come from the use of murine-derived feeder cells, xenogeneic matrices, or fromanimal-derived components found in the cell culture medium used to isolate or expand the stem cells. One of
the major challenges for the clinical use of stem cells is the exposure to undefined animal-derived products
during derivation and expansion of the cells. In this study, we have developed a xeno-free cell culture systemthat will support both hESC and hiPS cell systems. We have recently reported three new hESC lines(SEES1-3) which were derived and maintained on irradiated human mesenchymal stromal cells grown in
xeno-free culture media and substrate. SEES1-3 lines did not express non-human sialic acid reported as an
association with increased immunogenicity. Here we describe a xeno-free cell culture system that will support
both hESC and hiPS cell systems. In this presentation we will discuss our experiences in maintaining
pluripotent stems cells using these xeno-free media systems. Our center is officially approved hESC
derivation study by the Minister of Education and Science of Japan, and has built up the stem cell technology
for stem cell based therapy.
◆Biosketch
Hidenori Akutsu, MD/PhD
Chief, Division of Stem Cell and Reproduction, Department of Reproductive Biology,
National Research Institute for Child Health and Development
Dr Hidenori Akutsu became interested in nuclear reprogramming in mammalian species when he was a
research fellow at University of Hawaii under Dr Ryuzo Yanagimachi. This interest endured and motivated
him to undertake further research for epigenetic and nuclear reprogramming under Dr Minoru Ko at NIA/NIH
and Dr Kevin Eggan at Harvard University. While at Harvard University he also became an important part of
Dr Douglas Melton‘s team, deriving human embryonic stem cell lines which were later offered freely to thescientific community to facilitate the efforts of other scientists. His special interests are deriving humanembryonic stem cells for applying to clinical use, and egg development and epigenetic reprogramming.
Selected Publications:
1. X-Chromosome inactivation in cloned mouse embryos. Eggan K, Akutsu H, Hochedlinger K, Rideout W

214

●Concurrent Symposium C-23.1

3rd, Yanagimachi R, Jaenisch R. Science 290(5496):1578-1581, 2000
2. Maintenance of pluripotency and self-renewal ability of mouse embryonic stem cells in the absence oftetraspanin CD9. Akutsu H, Miura T, Machida M, Birumachi JI, Hamada A, Yamada M, Sullivan S,
Miyado K,Umezawa A. Differentiation 78(2-3):137-142, 2009
3. Aberrant silencing of imprinted genes on chromosome 12qF1 in mouse induced pluripotent stem cells.
Stadtfeld M, Apostolou E, Akutsu H, Fukuda A, Follett P, Natesan S, Kono T, Shioda T, Hochedlinger K.
Nature 465(7295):175-181, 2010
Contact Information:
2-10-1 Okura, Setagaya-ku,
Tokyo, 157-8535, Japan215

●Concurrent Symposium C-23.3

Development of pluripotent stem cells to female germ cells
Hsin-Fu Chen, Hong-Nerng Ho
National Taiwan University Hospital, Dept. of OB/GYN, Taipei, Taiwan
Introduction: Human embryonic stem cells (hESCs) and induced pluripotent stem
cells (hiPSCs) are unique stem cells characterized by the great capability ofunlimited self-renewal and under appropriate environments can develop into all cell lineages including germ
cells in human. These cells therefore will be useful as models for toxicology and pharmacology and also inclinical practice as sources of cell-based therapy for those patients suffering infertility, premature ovarian
failure (POF) or recurrent abortion, due to defects in germ cell quality and quantity. Recent data suggested
that ESCs can develop to early germ cells, but mature female germ cells remains a big challenge. Our Lab has
previously showed that hESCs were capable of differentiating spontaneously in vitro into cells expressing
germ cell markers and ovarian follicle-like structures. Recently we also derived a POF patient-specific hiPS
cells line. Therefore by using these stem cells as starting materials, germ cells hopefully may be produced for
future application.
Materials and Methods: We have created a reporter Oct4-eGFP hESCs line for tracking and enrichment ofgerm cells during in vivo and in vitro development. We differentiated NTU1 hESCs and H9 Oct4-eGFP
hESCs in vitro using a number of niche environments, including laminin-coated dishes, human granulosa cell
co-culture or conditioned medium (CM), mouse ovarian stromal cell co-culture or CM, and addition of growth
factors (retinoid acid, SCF and BMP4).
Results: We showed that granulosa cells co-culture and its CM increased the percentages of cells expressing
early germ cell marker SSEA1 (SSEA1+)(up to 40%) and/or Oct4-eGFP (Oct4-eGFP+) compared to laminin
alone (18%). Though the use of Oct4-eGFP line was not absolutely necessary, manual collected Oct4-eGFP+
cells on day 14 of differentiation significantly up-regulated specific germ cell gene VASA and late marker
GDF9 expression, although meiosis markers SCP1 and SCP3 and mature germ cell marker ZP3 were not.
These Oct4-eGFP+ cells developed into ovarian follicle-like structures after extended culture for 28 days. In
addition, supplement of retinoid acid also enhanced VASA and GDF9 expression in the differentiated cells,
whereas SCF and BMP4 did not. In addition, the injection of undifferentiated OCT4-eGFP hESCs into SCID
mice produced teratomas containing differentiated Oct4-eGFP+ cells. These eGFP+ cells and/or theirsurrounding cells also expressed early germ cell markers. Taken together, early human germ cells can be
identified and isolated through the use of pluripotent stem cells but can be enhanced by the use of transgenic
OCT4-eGFP hESCs reporter.
Conclusions: It is concluded that granulosa cells co-culture and CM may increase the percentages of cells
showing germ cell potential. SSEA1 alone may not be a good marker for isolating primordial germ cells fromdifferentiated hESCs, except perhaps at an early stage of differentiation. On the contrary, GDF9 may becomea useful marker for selection at later stage (after day 21). We propose that the protocol of differentiating hES
cells in granulosa cell co-culture/conditioned medium and/or retinoid acid supplement, followed by early
manual selection of Oct4-eGFP+ cells, and finally by selection with VASA and/or GDF9 (especially) markers
can become an effective strategy to enrich the cells with greatest potential for germ cell formation.
◆Biosketch(For Dr. Chen‘s Biosketch, please refer to his abstract for Concurrent Symposium C-19.4.)

216

●Concurrent Symposium C-23.4

Testicular stem cells as sources of fertility preservation and cell therapy
Kwang Yul Cha
CHA Health Systems and CHA University of College of Medicine, Seoul,
Korea
It is well known that many types of stem cells and somatic cells exist and are sustainedin testis tissue for the life of a male. At first, spermatogonial stem cells (malegerm-line stem cells, GSCs) self-renew and produce large numbers of differentiating germ cells that become
spermatozoa throughout the adult life. Specific models for in vitro proliferation and expansion of GSCs fromneonatal or adult testis have been developed by several researchers, and that has become a useful tool forstudying spermatogenesis mechanisms, and has important implications for the development of newtechnologies in transgenesis or medicine.
Recently, multipotent SSCs were derived from SSC in vitro and were proposed as a potentially useful sourceof pluripotent cells for cell-based therapy. This type of human cell may provide simple and non-controversial
access to individual cell based therapies without any ethical issues, but their efficiency for establishingheterogeneous testicular cell population was still extremely low and not well characterized. Last one istesticular mesenchymal stem cells (MSCs) with high proliferative activity. The MSCs were easily acquired bysimple culture and MACS sorting from male donors with either normal or abnormal testis physiology andhave shown similar growth kinetics, expansion rates, clonogenic capacity and differentiation potential as
MSCs. Further research and application of these types of testicular stem cells may allow individual cell-based
therapy without the ethical and immunological problems as well as infertility treatment.
◆Biosketch
Dr. Kwang Yul Cha is the Founder and Chancellor of CHA HEALTH SYSTEMS and CHA University inSeoul, Korea. Dr. Cha received his medical degree from the prestigious Yonsei University graduating summa
cum laude and performed his postdoctoral fellowship in endocrinology and infertility at the University ofSouthern California. Dr. Cha also served as a visiting professor at Columbia University where he establishedand operated CHA Columbia Fertility Center in collaboration with Columbia Presbyterian Hospital.
An internationally renowned medical expert and one of the most recognized authorities on reproductive
endocrinology and infertility, Dr. Cha pioneered the world's first successful pregnancy and birth through invitro maturation from immature oocytes and also the world's first successful birth with frozen oocytes
utilizing the technique of oocyte vitrification using grid.

217

●Concurrent Symposium C-23.5

In vitro production of functional sperm in cultured neonatal mousetestesTakehiko Ogawa
Yokohama City University
The whole process of spermatogenesis, which produce sperm from spermatogonia
through meiosis, has never been reproduced in vitro in mammals, nor in any otherspecies with a very few exceptions in some particular types of fish. In our effort to develop in vitro system for
spermatogenesis, we evaluated organ culture method to induce spermatogenesis in the testes of immature mice.
To make evaluation simple and easy, we exploited 2 lines of transgenic mice, Gsg2/Haspin-GFP and Acr-GFP,
which carry marker GFPs specific for meiosis and haploid cells. In our culture condition using serum-free
media, both Acr-GFP and Gsg2-GFP showed their expression mostly at the same time as they did in vivo andspermatids and sperm were produced from gonocytes or primitive spermatogonia. The spermatogenesis wasmaintained over 2 months in tissue fragments positioned at the gas-liquid interphase. The obtained spermatids
and sperm resulted in healthy offspring through microinsemination. In addition, neonatal testis tissues were
cryopreserved and, after thawing, showed complete spermatogenesis in vitro. Our organ culture method could
be applicable through further refinements to a variety of mammalian species, which will serve as platform for
future clinical application as well as mechanistic understanding of spermatogenesis.
◆Biosketch
Takehiko Ogawa, MD, PhD
1989:
Yokohama City University Graduate School of Medicine, finished MD course majoring at
Pathology.
1989-1995: Working as urologist at several hospitals including Yokohama City University Hospital.
1995-1998: Post-doctoral fellow at University of Pennsylvania, Veterinary School of Medicine (Ralph L.
Brinster's lab).
1998-present: Department of Urology, Yokohama City University School of Medicine.

218

STGO SESSIONISF SESSION
APART SESSION

219

●STGO Session STGO (1) 1

How to manage hyper- response to COS in IVF
Youssef Boutaleb
MoroccoOvarian hyper stimulation is the most serious complication of ovulation inductors
from a medical and medico-legal standpoint.
These complications present themselves as different clinical aspects.
There may be a pelvic accident such as a very high increase in the volume of the ovaries and the twisting or
rupture thereof. Or there might be phenomena altering several vital functions classified by Golan into fivestages:1- Severe abdominal distension
2- Nausea, vomiting, diarrhea, ovary 5 to 12 cm
3- As cites and/or hydrothorax with respiratory difficulties
4- Disturbance Coagulation and/or of the kidney function.
5- Abnormalities of coagulation and/or renal function .
The condition of these patients may become complicated and show evidence of:a state of shock, respiratory distress, disturbance of liver function, thromboembolic accidents, sometimes
leading to death. In certain instances sequels can occur: hemiplegic, amputation of members, ovariectomy.
Prevention remains the most effective treatment.
In other instances, the ovarian stimulation must be stopped. Sometimes just rest is sufficient. At other times
hospitalization is necessary in order to provide the patient with vascular expansion, punction of peritoneal or
pleural effusions to give relief to the patient, puncture of ovarian cysts - but rarely surgery. The use of anti
coagulants should be considered with care, as well as the use of anti-inflammatory substances, antihistamines,
inhibitors or conversion enzymes. Termination of pregnancy is considered a final recourse◆Biosketch
Doctor Youssef Boutaleb
Professor of Gynecology and Obstetrics
Formerly Medical Chief of Staff in Gynecology and Obstetrics of University Hospital Ibnou Rochd, Casablanca,
MoroccoFormer President, Moroccan Foundation of Medical Research
Former President, Moroccan Association for the Fight Against Cancer
Former President, Royal Moroccan Society of Gynecology and Obstetrics
Founder and Former President of the Moroccan Society of Fertility - Contraception
Former President of the World Congress I.F.F.S, 1989 Marrakech, Morocco
Author of Sterility of the Couple in the Arabo-Muslim Context
Royal Decoration as "Chevalier"220

●STGO Session STGO (1) 2

How to manage poor responders?
R. Frydman1,4, F. Lamazou1,4, V. Gallot1,4, M. Grynberg1,4,
R. Fanchin1,4, L. Hesters2,4, N. Frydman2,4, J. Taieb3,4
1
Univ Paris-Sud, Clamart, F-92140 ; AP-HP, Service deGynécologie-Obstétrique et Médecine de la Reproduction, Hôpital Antoine
Béclère, Clamart, F-92141
2
AP-HP, Service d‘Histologie-Embryologie Cytogénétique, Hôpital Antoine Béclère, Clamart,
F-92141 ; Univ Paris-Sud, Clamart, F-92140
3AP-HP, Service de Biochimie et Hormonologie, Hôpital Antoine Béclère, Clamart, F-92141
4
INSERM, U782, Clamart, F-92140
AMH is considered like a major prognostic factor of ovarian response during controlled ovarian
hyperstimulation. IVF in modified natural cycle has gained attention recently, even for both normal and poor
responder. We conducted a retrospective study on 415 cycles of IVF in modified cycles with an objective toevaluate the impact of serum AMH concentration on day 3 on the outcomes. Patients of less than 37 years ofage have been regrouped by AMH concentration: 0.04-0.49 ng/ml, 0.5-0.99 ng/ml, 1-1.49 ng/ml, 1.5-1.99
ng/ml, 2-2.249 ng/ml and 1.5-5 ng/ml. No statistically differences have been observed on cancellation rate,
failure of oocyte retrievals, embryo transfers rate, pregnancy or implantation rate. The embryo quality basedon the morphology on day 2 was similar in all the groups. In conclusion serum AMH concentration in day 3
does not seem to be correlated to pregnancy rate nor implantation rate, nor embryo morphology in IVF inmodified cycle for patients of less than 37 years of age. IVF in modified cycle should be proposed in firstintention to the patients with a poor prognosis characterized by a low AMH concentration inferior to 0.5 ng/ml
with an implantation rate of 27.18%.
◆Biosketch
Name:
Rene FRYDMAN
Birth date: November 3, 1943
Birth place: Soumoulou, France
Nationality: French
Current Positions and Rank: Professor, Faculty of Medicine, University Paris XI and Head of the
Department of Gynecology and Obstetrics of Antoine Beclere Hospital, since 1990 to 2010 – Head of MotherChild Department 2000-2010.
Awards of Distinction:
Officer of the French Legion d‘Honneur
Areas of Special Interest and Accomplishments:
My special areas of interest in Gynecology and Obstetrics include infertility and high- risk pregnancy. My
work in infertility led the first baby born in France as a result of in vitro fertilization in 1982, after embryo
freezing in 1986, after PGD in 2000, after IVM in 2003, after oocyte cryopreservation 2010, after PGD with
HLA matching in order to bring about the birth of a saviour sibling in January 2011.
My other area of interest has been biomedical ethics. My work in this realm led to many invitations to debate
the moral issues created by the use of the techniques of artificial procreation. I have had an active
participation in the preparation on the law on bioethics.

221

●STGO Session STGO (1) 3

The thin endometrium in ART: What to do?
Amina Oumeziane
TIZIRI: IVF center, Algiers, Algeria
The thin endometrium stills an unresolved problem in infertility. The negative
impact of thin endometrium in the pregnancy rate and live birth rate is still
controversial. In addition to the thicknesses of the endometrium, different studies
show that other parameters must be considered to evaluate the unresponsiveness of the the endometrium; theseinclude the following:

The volume of endometrium by 3D sonography the day of HCG.



The endometrium vascular patternThe immunological factors



The genomic analysis of the endometrium responsiveness

Different treatments have been suggested for improving IVF outcome. Oestradiol at luteal phase, Triptoreline
at luteal phase, Vitamin E, L.Arginine, Sildenafil, Low dose Aspirin
And recently, advance point to the role of cytokines in the proliferation of the endometrium by the use of
intrauterine infusion of G-CSF which may be considered as a new way of managing thin endometrium◆Biosketch
Dr AMINA OUMEZIANE: Gynecologist specializing in reproductive medicine. Currently: Medical Director
of TIZIRI: IVF center at Algiers
Fellowship at Cochin Hospital University Paris V France in 2000

222

●STGO Session STGO (2) 1

How to treat the female causes of repeated implantation failure?
Khaled Terras
Centre of Reproductive Medicine and Prenatal Diagnosis, Tunis, Tunisia
Implantation is a complex initial step in the establishment of a successful pregnancy.
Although embryo quality is an important determinant of implantation, temporally
coordinated differentiation of endometrial cells to attain uterine receptivity and a synchronized dialog between
maternal and embryonic tissues are crucial. The exact mechanism of implantation failure is still poorly
understood.
The causes for repeated implantation failure (RIF) may be because of reduced endometrial receptivity,
embryonic defects or multifactorial causes.
Various uterine pathologies, such as thin endometrium, altered expression of adhesive molecules andimmunological factors, may decrease endometrial receptivity, whereas genetic abnormalities of the male orfemale, sperm defects, embryonic aneuploidy or zona hardening are among the embryonic reasons for failureof implantation.
In this mini review, we discuss the suggested methods for evaluation and treatment of RIF: repeatedhysteroscopy, myomectomy, endometrial stimulation, immunotherapy, preimplantation genetic screening(PGS), assisted hatching, zygote intra-Fallopian transfer (ZIFT), co-culture, blastocyst transfer, cytoplasmictransfer, tailoring stimulation protocols and salpingectomy for hydrosalpinges.
Optimizing endometrial receptivity in fertility treatment will improve success rates. Evaluation of
implantation markers may help to predict pregnancy outcome and detect occult implantation deficiency.
Treating the underlying gynecological disease with medical or surgical interventions is the optimal current
therapy. Manipulating the expression of key endometrial genes with gene or stem cell-based therapies may
some day be used to further improve implantation rates.
◆Biosketch
Pr Khaled Terras is born in Bizerte (Tunisia) in January 1st, 1964
Medical studies, gynaecology and obstetrics degrees: faculty of medicine, Tunis
Diploma in the reproductive health, in foetal medicine and ultrasound in Paris universities (V &XIII).
Training in these fields was achieved in IVF unit of Baudeloque clinic in Paris and Harris birth right unit of
foetal medicine (Pr K. Nicolaides, London). He was a senior consultant at Charles Nicolle Hospital in Tunis.
Dr Terras is now member of the team of the ―centre of reproductive medicine and prenatal diagnosis‖ in Tunis.
He is the former general secretary of the Tunisian society of gynaecology and obstetrics, and the former
president of the scientific committee of the Arab Association of gynaecology and obstetric223

●STGO Session STGO (2) 2

How to manage sperm investigation after ICSI Failure
Moncef Benkhalifa1,2, Paul Cohen Bacrie2, Alain dalleac2,
Yves Menezo3
1
ATL R&D. Reproductive Biology & Genetics Laboratory. Paris
2
Eylau Laboratory/ Unilabs. Paris-Geneva
3
Dynabio Laboratories. Cherbourg France
Cytogenetics studies of infertile male karyotype revealed between 2 to 15% of abnormalites. In azoospermia
cases nearly 15 % are carrier of unbalanced karyotypes (mainly 47, XXY) but in oligospermia cases only 2 to
5% of chromosomes disorders were observed. More than numerical abnormalites translocations including
gonosomes are detected also. In the litterature its clearly reported that the male genome abnormalities can be
involved in oocyte activation failure, early embryo development blocking, implantation failure and pregnancyloss.
In male infertility several Y and autosomal linked genes are involved not only in testicular but also in ovariandevelopment. Others mutations in X chromosome (Kalleman gene, androgen receptor) or autosomal:(mytonic dystrophy, CFTR, 5-alpha -reductase Deficiency) are Known to interfere with normal
spermatogenesis . For male patient with abnormal caryotype the assessment of aneuploidy or chromosome
segregation in sperm is recommended prior IVF for genetics counselling.
During the spermatogenesis meiosis all chromosomes are equally submitted to the non disjunction with thenearly 10% of aneuploidy risk. Regarding reciprocal and robertsonian translocation the profile of segration isvariable from one translocation to another and between patient carrying the same translocation. For thisreason FISH analysis of sperm in such kind of cases is very informative to set up the strategy of couple
management (IVF, ICSI, Blastocyt, PGD).
Based on the profile of chromosome disjunction we will get an idea about the risk of embryo blocking,implantation failure and abortion. Another factor can contribute in ART failure more than chromosome
abnormalities is the DNA integrity and compaction of the entire male genome. An abortive spermatogenesis
or an oxidative stress can affect the DNA integrity by producing hight level of sperm DNA frangmentation.
We reported earlier that there is a relation between sperm parameter, the level of DFI and IVF outcomes.
During the spermiogenenis and germ cell final maturation, the efficiency of proteines replacement of histones
by protamines is important to keep the compaction of the genome and to protect the paternal transcripts which
involved in oocyte activation after fertilisation. Any unbalance of proteines replacement and /or between theration of protamine 1 and 2 can affect the competency of the spermatozoa for oocyte acticvation, embryo
development and implantation.
It‘s now established that mature spermatozoa contain a complex panel of m RNAS, studies performed earlier
showed that these mRNA have an important role in normal spermatogenesis and early embryo development .
From our experience using microarrays we showed that germ cells of oligospermic men have a majoralteration of gene expression profile when compared to normospermic individuals
In male to assure the final biological function of the spermatozoa any defect of the spermatogenesis(chromosome non disjunction, apoptosis and DNA fragmentation) and the spermiogenesis (proteines
replacement, compaction defect by denaturation, mitochondrial function or transcripts alterations) will play a
key role in IVF success mainly at embryo development and implantation failure levels.
◆Biosketch(For Dr, Benkhalifa‘s Biosketch please refer to his abstract for Concurrent Symposium C-16.2)

224

●STGO Session STGO (2) 3

How to improve laboratory procedures to enhance implantation?
Ghaya Merdassi
IVF Center, Hospital of Aziza Othmana, Tunis, Tunisia
The expansion of indications for ART has not only increased the number of patients
attending IVF clinics, but also the number of techniques currently in use. The
embryologist has a responsibility for the correct and justified application of ART in the laboratory. The
purpose of this report is to review the indicators and guidelines of good practice for each center of the ART toenhance the implantation rate. In fact, current methods of evaluation and quality measurement are reported as
the optimization of culture conditions, the operation of equipment and consumables. The application ofsystem quality management allows a high level of operation of the laboratory and the improvement of theimplantation rate through a better selection of gametes, zygotes and embryos.
◆Biosketch
Dr. Ghaya Merdassi
Date of Birth: 01/08/1976. Tunis
Marital status : not married
PRESENT ADDRESS:IVF Center, Hopsital of Aziza Othmana
Place de la kasbah Tunis, Tunisia 1008
EDUCATION AND TRAINING:
Inter University Degree: University of Versailles, Paris, France 2010
Master Degree: University of Claude Bernard Lyon, France 2
Biology Degree: University of pharmacy, Tunisia 2005
Pharma.D: University of pharmacy, Tunisia 2000
RESEARCH AREAS:
Cryopreservation of ovarian tissue
Cytogenetic analysis in teratozoospermia
Aneuploidy screening and polar body biops

225

●ISF Session ISF 2

Gonadotropin-releasing hormone agonist trigger: The way toeliminate OHSS
Arye Hurwitz
Professor, Head of the IVF Unit, Hadassah University Hospital,
Mount. Scopus, Jerusalem, Israel
After more than thirty years of IVF ovarian hyperstimulation syndrome (OHSS)
still remains a major threat to our patients in both long agonist and short antagonist cycles. It was firstsuggested by Itskovitz et al in 1988 that GnRH agonist (Buserilin) can induce the LH surge and oocyte
maturation while eliminating OHSS in 2 women who previously developed severe OHSS. Only in the last 5-6
years many centers have started using GnRH agonist to trigger ovulation in high risk and donor patients inorder to avoid OHSS. It was found that in 16 publications between 2006 - 2010 comparing hCG to GnRH
agonists there was not a single women who developed OHSS out of 2005 patients after GnRH agonist
triggering, while 92 out of 1810 (5.1%) developed OHSS after hCG. Even in GnRH antagonist cycles there
was an incidence of 2% of severe OHSS. It is suspected that GnRH agonists prevents OHSS by inducing
luteolysis of the corpus Luteum. This assumption is based on the studies by Nevo et al who demonstrated asignificant reduction of Estradiol and Progesterone as well as Inhibin A and Pro alphaC, all markers of luteal
demise. Since triggering ovulation with GnRH agonist causes a quick and irreversible luteolysis adequate
luteal support is crucial. When conventional luteal support was given (Uterogestan 200mg tid and Estrofem
2mg bid) the pregnancy rates were extremely low. However when the luteal phase was inforced by either 50
mg Progesterone I.M and 2mg Estrofem tid, or the 1500 IU hCG (one dose), from the day of oocyte retrieval
the pregnancy rates improved significantly and were similar to those obtained after hCG triggering. In
conclusion, we believe that OHSS should be a disease of the past and that GnRH agonist triggering should be
administered in high responders to totally abolish its occurrence.
◆Biosketch
Graduate of the Tel Aviv University Medical School.
Completed his residency in Obstetrics and Gynecology at Hadassah University Hospital at Mount Scopus,
Jerusalem.
Did his subspecialty in Reproductive Medicine at the University of Maryland in Baltimore, Maryland, USA.
Is currently the Director of the IVF unit at Hadassah University Hospital, Mount Scopus, Jerusalem.
Published over 100 articles in leading international journals.

226

●ISF Session ISF 3

From endometrial injury to IMSI: New approaches for repeated IVF
failureMartha Dirnfeld
Reproductive Endocrinology and IVF at the Carmel Medical Centre,
Faculty of Medicine, Technion - Israel Institute of Technology, Haifa,
IsraelIn vitro fertilization (IVF) has become a widely accepted clinical practice to assist reproduction. However,despite improved technologies and better results in recent years, the majority of patients do not have a
successful pregnancy following a single IVF treatment.
The availability of assisted reproductive technology (ART) and delivery rates per oocyte retrieval vary greatly
between countries. Per aspiration, pregnancy rates ranges are 17% to 42% and delivery rates 8% to 34%
Repeated treatments and failures have negative impact on quality of life. Each failed cycle incurs substantial
financial costs. The expense, time, stress, and frustration felt by couples and physicians has led to a search for
new drugs and technologies that will increase success rates. The aim of this presentation is to establish the roleof various therapies for patients with repeated implantation failures.
Although some techniques have shown to increase pregnancy rates in women with poor prognosis due tospecific conditions such as Hydrosalpinx, intrauterine polyps or adhesions, specific endocrine diseases orlong-term gonadotrophin releasing hormone agonists for women with endometriosis, many infertile women
still fail to conceive, despite repeated transfers of high-quality embryos.
A number of strategies have been suggested to improve unexplained repeated implantation failures. Various
techniques in the IVF laboratory have been used on sperm oocytes and embryos in conjunction with IVF to
increase the pregnancy rate. Empiric drugs before after or adjacent to ovarian stimulation have been used andvarious procedures have been advised to patients with repeated implantation failures .
Although mechanisms of actions have been proposed, justification for the use of some therapies is usuallyempirical, and based on physicians‘ personal views. Controlled randomized studies of adjuvant therapies inwomen who have had repeated IVF failure are often difficult to perform, due to the great variation between
patients and the large sample size required.
The main factors found to influence the outcome of IVF and intracytoplasmic sperm injection (ICSI) include
maternal age, the number of oocytes retrieved, sperm quality, the number and quality of the embryos
transferred, the technique and ease of embryo transfer (ET), and endometrial receptivity. All the above should
be addressed in patients with repeated failures.
Individual review of each implantation failure and the use of some adjuvant therapy may improve the outcomeof IVF.
Review of the literature shows that none of the available adjuvant therapies has a clear advantage to all
patients. Notably, adjuvant therapies have been administered without a diagnosis as to whether the failure toconceive is due to a maternal or fetal factor. Salpingetomy in cases of Hydersalpinx and conservative
Myomectomy of fibroids encroaching the cavity may improve implantation in a subgroup of patients. Highmagnification (over ×6000) provides the identification of spermatozoa with a normal nucleus and nuclear
content. Injection of spermatozoa selected according to fine nuclear morphology under high magnification
may improve the clinical outcome in cases with severe male factor infertility. However, Intracytoplasmic
morphologically selected sperm injection (IMSI) did not provide a significant improvement in the clinicaloutcome compared with ICSI.
If the embryos are genetically abnormal, no maternal adjuvant treatment will improve the pregnancy rate, andthe genetic aberration will confound the results. Similarly, PGS will not be effective if implantation failure

227

●ISF Session ISF 3

is due to a maternal factor. Therefore, some of the therapies that have not been confirmed may prove
efficacious in subgroups of patients.
Even when failure to conceive originates in the woman, certain adjuvant therapies may benefit only womenwith particular characteristics. As examples, endometrial biopsy or minor injury to the endometrium (Pipel)
may benefit patients with a thin or nonresponsive endometrium
Uncertainty of the effectiveness of sildenafil may be due to the confoundingof fetal factors. Similarly, heparin may be effective against antiphospholipid antibodies other than lupus
anticoagulant or anticardiolipin antibody. Profilaxis antibiotics may be suitable for some patients.
Many patients with repeated failure are also poor responders. Treatments such as Steroids , Growth Hormone
and DHEA will be discussed. Presently, the diagnosis of IVF failure is not sufficiently specific to indicate
definite adjuvant therapy.
In light of patients‘ easy access to updated articles, physicians need to be especially prepared to answer
questions raised by couples that have repeatedly failed to conceive. Physicians endeavor to provide treatmentthat may be beneficial; as their role is not only to withhold treatment until sufficient randomized trials are
conducted. Treatment often needs to be ―tailor-made‖ to suit the individual patient.
◆Biosketch
Prof Martha Dirnfeld
Graduated medicine at the Technion, Israel, Institute of Technology, Haifa and Board certified in OB/GYN at
the Carmel medical centre.
Since 1986 Head of Division - Reproductive Endocrinology and IVF at The Carmel Medical Centre, Facultyof Medicine, Technion - Israel Institute of Technology, Haifa, Israel.
President of the Israeli Fertility Society (IFS) and Israeli representative to ESHRE – National Committee
Representative, (NRC) .
Worked in leading IVF Centres in Europe.
During 3 years (1982-1984) in Belgium with IVF team at the Free University (ULB) and Honorary Senior
Clinical Research fellow at Bart's and the London Queen Mary's School of Medicine and Dentistry, London
In 2002-2004 invited to be General Manager at London Bridge Fertility-IVF Gynecology and Genetic Centre.
UK Leading center in ART and PGD and since then regularly visits for consultations at the same centre.
Prof M.Dirnfeld Research interests include:•••

Various research projects in Reproductive endocrinology and assisted reproductive technologies
Oocyte and Embryonic microenvironment
Factors involved in Implantation mechanisms and implantation. .

Published more than 150 papers, reviews and Abstracts at international conferences and Chapters in 6 books.
Invited speaker at national and International Conferences.

228

●ISF Session ISF 4

High ART success rates when financial constraints are completely lifted
Seidman DS, Lande Y, Maman E, Baum M, Dor J, Hourvitz A.
IVF Unit, Department of Obstetrics and Gynecology of the Chaim Sheba
Medical Center, affiliated to the Sackler Faculty of Medicine, Tel Aviv
University, Tel Aviv, Israel
Israel has a unique situation because IVF treatments are fully covered by the national
health insurance and thus are provided free of charge to all women until their first two children are born.
Israeli patients are offered easy access to IVF clinics where they can immediately undergo, when medicallyindicated, an unlimited number of IVF treatment cycles. We assumed that for infertile patients the chances of
becoming pregnant and having a live child should be greater in Israel than in most other countries because thefinancial cost does not play a role in the patient‘s decision to obtain treatment.
We undertook an historical prospective cohort study that included all couples referred to the Sheba Medical
Center IVF unit who met the following inclusion criteria: [1] women aged <35 years at the time of referral, [2]women with primary infertility, [3] women with no IVF treatments before referral, [4] first IVF cycle
performed between 2001 and 2002, and [5] a minimum of 12 months‘ infertility before initiation of IVF
treatment.
During the 5- to 7-year follow-up period, 95.5% of couples conceived, and 89.6% of couples gave birth to alive infant. Of these couples, 81.3% achieved a live birth within the first 4 years of the follow-up period, and85.1% within eight treatment cycles. Of the 14 couples (10.4%) who did not give birth to a live infant, five
adopted, two divorced, four are still undergoing IVF treatments, and three (1.8%) decided not to become
parents.
We concluded that young couples beginning IVF treatment in an environment free of economic hurdles can bereassured that they have an excellent chance (~90%) of achieving a live birth within 4 years. When IVF isprovided free of cost, very few couples discontinue treatment before a live birth is achieved.
◆Biosketch
Daniel S. Seidman, MD, MCSc
(For Prof. Seidman‘s Biosketch please refer to his abstract for Concurrent Symposium C-20.3)

229

●ISF Session ISF 5

Autologous transplantation of very thin ovarian fragments which
preserve the ovary's main cortex structure lead to successful
pregnancy
Ariel Revel1, Meital Lebovich1, Alex Simon1, Neri Laufer1, Einat
Eizenmann1, Eduard Mitrani2
1
Dept. of Obstetrics and Gynecology, Hadassah University hospital,
Jerusalem, Israel
2
The Alexander Silberman Institute of Life Sciences, Hebrew university, Jerusalem, Israel
The improved survival rates among patients with hematological malignancies, such as lymphoma andleukemia, are shifting areas of focus towards understanding and preventing treatment-induced sequel. Of these,
infertility is one of the most devastating consequences for patients with reproductive potential. The degree oftreatment-induced gonadal dysfunction depends on age and gender-related differences, the type and dosage ofchemotherapy used and the field and cumulative dose of abdomino-pelvic irradiation. There is also theinteresting phenomenon of reduced pre-treatment fertility among male lymphoma patients. At present, theonly established methods of fertility preservation are cryopreservation of sperm, oocytes and embryos, as wellas gonadal shielding and transposition of ovaries during irradiation. Several other methods, such as
cryopreservation and subsequent transplantation of gonadal tissue and the gonadoprotective role of hormonal
suppression, are under investigation. Pre-pubertal patients present a unique constellation of fertility
considerations, especially as embryo and sperm cryopreservation are not applicable to this age group. We will
present our data on ovarian cryopreservation for the past decade for various indications. We will detail our
19-year-old thalassemic patient who had tissue from one of her ovaries cryopreserved prior to bone marrowtransplantation, total body irradiation and sterilizing chemotherapy. As expected, premature ovarian failure
resulted from this treatment. Transplantation of her thawed ovarian tissue resulted in return of menstrual
cycling and the patient then underwent several IVF cycles. The patient, however, had poor ovarian response to
hyperstimulation. We thus considered an alternative approach based on the observation that very thin ovarianfragments that preserve the basic ovarian structure [ovarian micro-organs (MOs)] induce angiogenesis andremained viable after autologous transplantation in animals. Preparation of autologous tiny ovarian fragments(MO)s and re-implantation into our patient resulted in IVF pregnancy and delivery of a healthy baby. The
presentations will discuss the state of the art in the field of fertility preservation before gonadotoxic
chemotherapy.
◆Biosketch
Born in Strasbourg, France (1961), Married (Nili, 1984) and father of four. MD from Hebrew University(1988). Israel board certified in Ob/Gyn following residency at Hadassah Mt. Scopus (1996). Completed
fellowship in reproductive medicine at the Department of Obstetrics and Gynecology, University of Toronto,
Ontario, Canada (1998-2000). Currently, staff physician, in vitro fertilization (IVF) unit and Department of
Obstetrics and Gynecology, Hadassah. Member, Endocrinology circle, Hebrew University Medical School.
Manager of the oocyte donation (OD) program. Leader of the fertility preservation program. Clinical andresearch fields include IVF, embryo implantation, endometriosis, polycystic ovarian syndrome (PCOS),
laparoscopic and hysteroscopic surgery and ovarian cryopreservation. Served as treasurer of the Israel Fertility
Association (IFA) executive committee (2004-7). Board member Special Interest Group for Endometriosis &Endometrium (SIGEE) of the European Society for Human Reproduction and Embryology (ESHRE), Boardof directors for the cancer institute for fertility preservation. Hobbies include snowboarding, tennis and movie
scriptwriting.
Specialties: infertility, oocyte donation, ovarian cryopreservation and transplantation, in vitro fertilization,
endometriosis, polycystic ovarian syndrome, fibroids, operative hysteroscopy, laparoscopic surgery

230

●ISF Session ISF 6

How should we consult candidates for fertility preservation due to amalignant disease regarding their expected ovarian response to COH?
Shevach Friedler
Sasckler Faculty of Medicine, Tel Aviv University, Israel
The Infertility and In-Vitro Fertilization Unit, at Assaf-Harofeh Medical
Center, Israel
Evidence based consultation is crucial for candidates for fertility preservation facing chemo/radiotherapy due
to a malignant disease, regarding their expected ovarian performance after COH. A systematic MEDLINE(Pub-Med) search revealed a total of only seven retrospective, case control studies matching our inclusion
criteria. A meta-analysis was performed based only on published results in which the outcome of thesepatients was compared to age matched healthy patients undergoing COH for IVF/ICSI. The available
literature so far includes patients with variable age groups and the outcome is not stratified according to thespecific malignant diseases. COH protocols used are variable, the majority using GnRH antagonist. However,
it is evident from this meta-analysis that candidates for fertility preservation with a malignant disease, reacheda significantly lower ovarian response compared with their healthy controls. Their fertilization rates werecomparable. The number of cryopreserved oocytes or embryos is not stated systematically in the studies
included in this review. Since only a minority of the embryos underwent thawing and transfer the data
available does not allow to draw conclusions regarding the true reproductive potential of fertility preservationin these patients.
◆Biosketch
Prof. Shevach Friedler is an associate professor at the Sackler School of Medicine, Tel-Aviv University and asenior physician at the Infertility and In-Vitro Fertilization Unit, at Assaf-Harofeh Medical Center, Israel. He
is also the director of the male and female infertility clinic of Kupat Cholim Meuchedet, Tel-Aviv. He
obtained his M.D.degree at the Medical faculty of the Hebrew University of Jerusalem. Following
specialization in Obstetrics and Gynecology in Jerusalem, he also completed a fellowship in Reproductive
Endocrinology at Stanford University, California, USA. For the last twenty six years he works in the field of
infertility and in vitro fertilization, currently in a university affiliated IVF unit, treating over 1000 IVF/ICSI
cycles per year. He has authored over 80 publications including original articles, case reports, chapters inbooks and more, dealing with various aspects of infertility treatments, hysteroscopy, several pioneering papers
concerning surgical sperm retrieval in azoospermic patients as well as different treatment modalities inin-vitro fertilization.

231

●APART Session: APART 1

Clinical efficacy of adipose MSCs in human
Byung-Rok Do
HurimBioCell Inc., Seoul, Korea
Stem cells are unique cell populations with the ability to self-renewal anddifferentiation, and categorized into embryonic stem cells (ESCs), adult stem cells andinduced pluripotent stem (iPS) cells. Adult stem cells existed in most of functionally mature tissues, such asepithelia, blood, germ line, brain, muscle and pancreas and are known to contribute replenishment of tissues
after normal cellular senescence or injury. Mesenchymal stem cells (MSCs), one of the adult stem cells, areexcellent sources for developing a practical use in cellular therapeutics. They have no ethical problem andmany advantages which is multipotency, auto-transplantation and safety from carcinogenesis. However,heterogeneous populations of MSCs are inevitable consequence of isolation methods. It makes difficult todefine stem cells and elucidate therapeutic mechanisms. Recently, MSCs are defined as an ability of plastic
adherence, capacity of multipotency, and characteristic phenotype at the 2007 conference of the International
Society for Cellular Therapy (ISCT). The phenotype expression is CD73, CD90, and CD105 positive andCD34, CD45, CD14 or CD11b, CD19 or CD79a negative, and HLA-DR low.Adipose tissues, originatingfrom embryonic mesodermal tissues, contain MSCs named as adipose derived stem cells (ASCs) with thehighest frequency among mesenchymal stem cells about 3 x 104 ~ 2 x 106 cells/ml aspirated fat tissue. ASCs
show similar characteristics with bone marrow derived MSCs. They can be differentiate into adipocytes,
chondrocytes, osteocytes in vitro, and can also express of phenotypic characteristics of endothelial cells,
neural cells, smooth muscle cells, skeletal myoblasts, and cardiac myocytes after differentiation induction.
ASCs have regarded as a valuable source for regenerative medicine, aesthetic plastic surgery and oncologicaldisorders. Many researchers have investigated that ASCs promote regeneration of damaged tissues byangiogenic effect, paracrine effect and immune modulation through animal models such as stroke, myocardial
infarction, liver disease and diabetes. Especially, abilities of various cytokine secretions and immune
modulation of ASCs have expected a various application in pharmaceutical or cosmetic. Moreover, ASCs
have provided the important cues for cell survival in the damaged tissues during long term tissue repair, sothey may become major cellular candidates in attempts to clinical applications. Recently, attractive
investigations are in progress for functional improvement of genital glands and differentiation into germ celllines. Especially, germ cell differentiation of ESCs, iPS cells and oocyte differentiation of MSCs have
gradually extended into clinical field. In Summary, development of effective cell therapy using human ASCs
should be accomplished technical point such as ex vivo expansion, controlled differentiation, stable phenotype,genetic stability, optimal microenvironment conditions, and functional conservation after transplantation.
◆Biosketch
Name: Byung-Rok Do, Ph.D.
Date of Birth: Jan. 26. 1962.
EDUCATION
1995 - 1998 Department of Biology, Doctor Course of Graduate School, Hanyang University, Seoul, Korea
PROFESSIONAL POSITION
2005 - Director Research Institute, Hurim BioCell Inc. Plural Professor in Hanyang University
1997 - 2003 Infertility research center, KangSeo MizMedi Hospital.
1990 - 1997 Infertility medical center, CHA General Hospital
RESEARCH INTEREST: Stem cell isolation, cryopreservation and guided differentiation Growth,development of human fetal ovarian tissues and follicles in vitro Human oocyte maturation in vitro
BIBLIOGRAPHY International Journal (12), Korean Journal (15) Proceedings and Poster Presentation;
International (62), Local (40) PATENT and PATENT ON A NEW DEVICE; Patent - 15232

●APART Session: APART 2

Optimized liposuction method for mesenchymal stem cell extraction
and subsequent culture in lean female patients
Ken Nakama
Coeur clinic
Objective: It has been recently found that compared to other sources,
stromal-vascular cells = the progenitors of human fat tissue - are relatively easy to extract and can be
efficiently cultured into mesenchymal stem cells (MSC).This could become a valuable source of MSC withpotential applications in the field of gynecology. The present study evaluated the efficiency of an optimized
liposuction method using Ringer‘s lactate solution for cell extraction.
Materials and methods:
Conventionally when liposuction is applied to the lower abdomen the ―tumescent‖ method is used. Thismethod developed in 1980 requires the subcutaneous injection of a large amount of isotonic saline solution.
Although, currently this is the established method for weight reduction purposes it might be suboptimal for
MSCextraction. It has been found previously that compared to the currently used saline solution survival rates
were significantly higher when extracted stem cells were placed into Ringer‘s lactate solution. Anotherpotential problem related with MSC extraction is that in lean patients, sufficient subcutaneous fat tissue isoften lacking in the lower abdomen and the procedure has to be performed in other body areas.
Results:
Randomly selected gynecological outpatients who were candidates for liposuction and MSC extraction andculture had a mean age of 43.5 years and an average BMI of 20.9. In contrast, in a control sample of
two-hundred patients who were scheduled for a weight-reduction liposuction intervention, the mean age was
32.2 years and the average BMI reached 24.7 (statistically significantly different). Compared to isotonic saline
solution MSCrecovery rates were not significantly different when Ringer‘s lactate was used. The differencesin MSCrecovery rates from different body areas have to be evaluated in a future study.
Conclusions:Compared to the general liposuction patient population, gynecological outpatients who were candidates forliposuction and MSC extraction and culture were older. They had a lower BMI and diminished body fat
tissue. Optimized liposuction by modifying the currently established ―tumescent‖ method with the use of
Ringer‘s lactate and using alternative body areas in lean patients might improve the results of subsequent
MSC culture.
◆Biosketch
M.D.degree University of CungUng, Seoul 1987
Assistant Resident in Surgery, University of Catholic HospitalGent, Seoul, May1987-Aug1991
ICRC, République du Burundi, Sep1991-Aug1992
Fellow in the Department of Transplant Surgery, University Hospital, May1993-May1995
Fellow in the Department of Anesthesiology, Tokyo Med. University Hospital, Tokyo, May1995-May1999
Coeur clinic, Dec2009-present

233

●APART Session: APART 3

Vitrification of adipose MSCs
Noriko Kagawa
Advanced Medical Research Institute of Fertility, Kato Ladies Clinic,
Tokyo, Japan
Mesenchymal stem cells (MSCs) are capable of differentiation into a variety of cell
types including osteoblasts, chondrocytes and adipocytes and are a promising source for cell-based therapy inregenerative medicine. Stem cell population isolated from human adipose tissue contains MSCs and their
biological characteristics are similar to those derived from bone marrow. Since human adipose tissue can be
harvested in large quantity by a minimally invasive liposuction procedure, tissue cryopreservation system isrequired to keep their freshness. Here, we developed successful vitrification method for cryopreservation ofhuman adipose tissue. Human discarded adipose tissues were donated from anonymous patients who
underwent a liposuction procedure and signed our informed consent. Human adipose tissue were vitrified byusing human ovarian vitrification method (Cryotissue R: Kitazato BioPharma, Kagawa et. al., 2007) withmodification. After thawing vitrified tissue, MSCs were isolated and survival rate of MSCs were examined.
Survival rate of MSCs isolated from vitrified adipose tissue was 78.9% and there was no significant differencecompared with MSCs from fresh adipose tissue (83.4%). These results indicate that our new vitrification
method is safe and effective for cryopreservation of human adipose tissue. We speculate that our vitrification
method will open the door for feasibility to access samples to promote stem cell research and translational
medicine.
◆Biosketch
Dr. Noriko Kagawa obtained her Ph.D. in 2005 from the Kyoto University, Japan. She is currently the seniorscientist at the Advanced Medical Research Institute of Fertility in Kato Ladies' Clinic, Tokyo, Japan, theworld's largest human IVF unit. In 2000, she started animal embryology and studied Assisted Reproductive
Technologies, for example, IVM, IVF, vitrification, embryo culture in pig, and established the porcine
follicular growth system using SCID mice. She moved to the human IVF field in 2004, and has developed
vitrification methods for mammalian ovaries and ovarian tissues in mice, cat, dog, lion, giraffe, bovine, human.
Her major research interests have focused on in vitro growth and in vitro maturation for preantral follicles ofadult mammals as well as on vitrification of Germinal Vesicle stage oocytes and various organs for unlucky
patients.

234

●APART Session: APART 5

Preparation of Adipose-Derived Stem Cells
Rie Yamadera, Yuji Takehara, Chiaki Sano, Kazumi Kawashima,
Yoko Ito, Osamu Kato
Kato Ladies Clinic
Adipose-derived stem cells (ASCs) are increasingly drawing attention as a tool inregenerative medicine, because large volumes of adipose tissue can be collected from patients using aminimally invasive procedure.The general method for separating cells from adipose tissue involves the use of
enzymes. The cells separated from adipose tissue belong to heterogeneous cell populations, including
blood-derived cells, vascular endothelial cells, adipose precursor cells, and other cells. Recently, someresearch groups have reported the existence also of a very small number of stem cells, similar to mesenchymal
stem cells (MSCs), in the cell populations separated from adipose tissue. These cells are called
adipose-derived stem cells (ASCs). We performed flow-cytometric analysis and CFU-F (colony forming
units-fibroblast) assay to confirm the number of ASCs included in this adipose-derived cell populations, andobtained results that were consistent with previous reports.Furthermore, it is possible to isolate and expand thestem cells by adherent culture of the cells separated from adipose tissue; this represents purification of ASCs.
We confirmed a monogenous population of the cells under culture and also that the characteristics of the cells
did not change under post-culture conditions.In this session, we shall introduce a general protocol to separate
the cells from adipose tissue, and also the approach to culture that we employed to isolate the ASCs.
◆Biosketch
2007: Graduated from University of Tsukuba, Ibaraki, Japan, with B.Sc.
2007-Present: Kato Ladies Clinic
2009-2010: Visiting scientist, physiology, University of Keio
Laboratory medical technologist
Genetic counselor235

●APART Session: APART 5

Discussion about clinical applications of adipose MSCs
Yuji Takehara
Kato Ladies Clinic in Japan
Mesenchymal Stem Cells (MSCs) have recently received attention in clinical fields tocure serious illnesses for which no effective treatments exist, such as brain diseases, diabetes, hypertension,
joint pain due to cartilage erosion, and myocardial infarction. While there is hope for regenerative medicine
using stem cells, on the other hand, there is also anxiety about its effectiveness and side effects. Because wehave very limited knowledge about stem cells and its regulatory pathways, new reliable reports are awaited.
There are three types of stem cells that are potentially useful in the field of regenerative medicine. Embryonic
Stem (ES) cells were established by Dr. Martin Evans in 1981, and accumulated data has given humans bright
hope for cure of diseases, especially after the establishment of human ES cells by James Thomson in 1998.
Although scientists have clarified in detail the mechanisms of actions of ES cells and also their efficacy,
establishment of human ES cells involves sacrifice of blastocysts which have the potential to develop into
human beings, as the inner cell mass of certain blastocysts is the source of ES cells. Once an ES cell line isestablished, it can be used permanently. This is an attractive idea, however, we need to apprise the couple
donating their embryos to the basic scientist who will handle their precious blastocysts about this ethical issue.
Another problem of use of ES cells in therapeutics is that it may evoke a rejection response whenadministered to the patients, because the ES cells originate from a different human and the chance of HLA
type matching is very low. As a result, an ES cell banking system that will cover different types of HLAs
will be required, and many ES cell lines are needed, which would need many healthy blastocysts to be
destroyed. This ethical issue prompted scientists to establish iPS (induced Pluripotent Stem) cells, which canbe obtained from mature adult cells, such as skin. In this regard, the greatest success in the establishment of
iPS cells was reported by Dr. Yamanaka in 2006 and 2007, which again raised enthusiasm about curing
difficult-to-cure diseases. iPS cells are not derived from blastocysts, which can develop into human beings,
but from mature cells which are destined to be eliminated by cell death. No ethical disputes would be
provoked when these cells are collected from the skin of the patient himself/herself. Another advantage of iPS
cells is that administration of iPS cells is not associated with the risk of a rejection response, because thesecells originate from the patients themselves and the HLA type is exactly matched with that of the patients.
Despite these miraculous advantages, iPS cells have the potential to induce tumors as a result of induction ofviruses. This possibility raised fear in respect of the clinical application of these cells. At the time of discoveryof iPS cells, it was suggested that a two-year period would be needed to get safe iPS cells. This was not
related to the production time, but the time needed to check whether the iPS cells will induce production oftumors. Recent competition to avoid virus use in the production of iPS cells again raises hope of obtaining
safe iPS cells, but they still remain to be established. Perhaps iPS cells will also be useful for checking theeffectiveness or side effects of newly produced drugs before clinical application. On the other hand, adipose
tissue-derived MSCs (ADMSCs) are easy to obtain from abdominal fat tissue, and this liposuction procedure
is applied in the cosmetic field for the purpose of weight loss. MSCs have the potential to differentiate into
any type of cells required at the site of damage. There are some reports that the administration of MSCs bydrip infusion may sometimes be dangerous as they may be associated with the risk of pulmonary embolism.
Because cultured MSCs are larger than the original parent cells and there is a surface antigen whose function
is to interfere adhesion, and number of this antigen decreased after culturing MSCs, decrease in the expressionof surface antigens interfering with cell adhesion after culture of MSCs may increase cell-to-cell adhesion,
resulting in the formation of bigger cells. We submitted a clinical research proposal to the Japanese Healthand Labour ministry in September in 2010, because clinical research using large amounts of stem cells

236

●APART Session: APART 5

requires approval of the Japanese government. The government recommended that we perform animal
experiments and establish our own animal laboratory to conduct the experiments. Therefore, it is not easy tostart a clinical study in Japan because of the Japanese Government‘s regulations. We believe that MSCs are
the best candidate for regenerative medicine, because there are no related ethical issues and these cells areeasy to obtain and also easy to handle.
◆Biosketch
Yuji Takehara MD., Ph.D.
1983
Graduated from Keio University, School of Medicine, Tokyo, Japan
Received M.D.
1991
Assistant Professor, Department of Obstetrics and Gynecology, Tokyo Dental College, Chiba, Japan
1991
Research Fellow at Johns Hopkins University, School of Medicine, Baltimore, MD, U.S.A.
1992
Instructor at Johns Hopkins University, School of Medicine, Baltimore, MD, U.S.A.
2008
Vice director of Kato Ladies Clinic
2011
Visiting Professor of Kinki University
Director of KLC Regenerative Medical Laboratory

237

ORAL COMMUNICATIONS

238

●Oral Communications

O-001

Does male reproductive tract CD52 (mrt-CD52) prevent complement activation via
binding C1q?
○L. Hardiyanto, A. Hasegawa, S. Komori
Hyogo College of Medicine, Kobe, Japan[Introduction] Human CD52 is a glycosylphosphatidylinositol (GPI) anchored peptide expressed in lymphocytes aswell as the male reproductive tracts (mrt). mrt-CD52 is known to be a pathogenic antigen for immunologicalinfertility. A human monoclonal antibody (Mab H6-3C4) generated from an infertile woman has strong sperm
immobilizing activity with complement and specifically recognizes the N-linked carbohydrate of mrt-CD52 but not
lymphocyte CD52. In addition, mrt-CD52 interfered with the classical pathway but not lectin binding or alternative
pathways of the complement system. In this study we focused on the C1q molecule as a binding component to
mrt-CD52. [Material and methods] mrt-CD52 was purified from human seminal plasma or sperm membrane by
extraction with a mixture of chloroform: methanol: water. For examination of mrt-CD52 binding to C1q, we
carried out immunoprecipitation analysis by incubating a reaction mixture of mrt-CD52, human serum for C1q
source, and Mab H6-3C4. N-glycosidase F (NGF) and phosphatidylinositol-phospholipase C (PI-PLC) were used
to release the N-linked carbohydrate and the GPI-anchor from mrt-CD52, respectively. When NGF treatedmrt-CD52, the carbohydrate moiety was isolated by ConA sepharose resulting unbound fraction comprised
GPI-peptide portion and bound fraction comprised carbohydrate moiety, respectively. Those fractions then reactedwith human serum and Mab H6-3C4. The formed immunoprecipitate was analyzed by a western blotting with the
ECL method probed with anti C1q antibody. Sperm immobilization test (SIT) was carried to determine the effectof purified-CD52. [Results] Immunoprecipitate formed from the reaction mixture of mrt-CD52, human serum and
Mab H6-3C4. After treatment of mrt-CD52 with NGF, immunoprecipitate formed from the reaction mixture of
mrt-CD52, human serum and Mab H6-3C4. The immunoprecipitate was formed by the carbohydrate moiety
reaction with human serum and Mab H6-3C4 while GPI-anchor peptide was not. mrt-CD52 treated with PI-PLC
also formed an immunoprecipitate with human serum and Mab H6-3C4. The C1q molecule (29 kDa) was detectedin the formed immunoprecipitates. In SIT, purified-CD52 showed an inhibitory effect on sperm immobilization
value (SIV). [Conclusion] These results indicate that the carbohydrate moiety of mrt-CD52 binds to C1q but not
the GPI-linked peptide. mrt-CD52 may inhibit the complement activity induced by the sperm antigen-antibody
complex in female genital tracts.

O-002

Technology Research of Prostatic Urethral Irrigation and Drainage Catheter
Injection○Weidong Huang
Xinjiang Jiayin Hospital Center for Reproductive Medicine, Urumqi, China

Chronic prostatitis is a common and frequently-occurring male disease, it can seriously affect the semen quality.
Cause of its complexity, longer duration and easy to relapse, so the clinical efficacy is uncertain. On the basis of
the two-purse three-cavity prostatic urethra injection irrigation and drainage catheter, Professor Huang Weidong
developed the catheter patents technology: it directly against because of urinary reflux, urinary prostate smoothmuscle spasm, immune response, bacterial infection caused by factors such as the prostate duct obstruction andinflammatory exudate within the lumen of the prostate is difficult to discharge these two clinical problems, throughthe involvement of the prostatic urethra characteristics of catheter positioning section and thrombolytic drugs
containing antibiotics and mixed liquid composed of the formation of pressure washing simulation prostatitis occur
when the "urine reflux" type of "liquid reflux" to clear the prostate duct openings clear of inflammatory exudate
and the prostate gland to relieve the obstruction. Because of its direct, safe, effective, simple, painless, repeatable
and non-injury characteristics, making the cure rate of chronic prostatitis has been significantly improved.

239

●Oral Communications

O-003

Measurement of reactive oxygen species in neat and washed sperm, is there any
difference?
○Mohammadreza Moein, Nasim Tabibnejad, Jalal Ghasemzadeh
Yazd Research and Clinical Center for Infertility, IranObjectives: Reactive oxygen species (ROS) are regarded as a major cause of infertility especially in idiopathic
male infertility. ROS level have been measured both in washed and neat sperm. So in this study we determined thelevel of ROS in neat and washed samples of infertile men to see which sample could show more reliably the
overproduction of ROS and oxidative stress.Materials and Methods: we collected semen sample from 35 infertile
men referred to our andrology clinic. Patients with pyospermia and patients who had history of antioxidant
treatment were excluded from study. The ROS levels were assessed by a chemiluminescense assay, using Luminol
as a probe, with an Autolamat LB 935 luminometer (Berth Hold Technologies, Germany) before and after washing.
Semen parameters and ROS level were compared in neat and washed samples.Results: The mean age of subjects
were 31.43 years ranging from 23 to 60 years. Mean total sperm concentration in neat samples was 95.94
million/ml and mean quick and slow progressive motility was 17.82% and 31.14% respectively. Mean ROS levelin this samples was 216.34 RLU. The mean sperm concentration in washed samples was 57.57 million/ml which
was significantly lower than neat samples (p=0.000). The mean quick and slow progressive motility was 49.37%
and 28.91% respectively. The quick motility was significantly higher than neat samples with (p= 0.000), but slow
motility has not significant difference with neat samples (p=0.494).The mean ROS level was 62.06 RLU in washedsamples. The mean ROS level was significantly lower in washed sperm (p=0.01). Conclusion: washing sperm canaffect ROS level in semen samples of infertile me. Although sperm processing seems to increase the level of ROS
in semen samples, but removal of immature germ cell and white blood cells will decrease ROS level in seminal
plasma. Which one is more accurate for predicting oxidative stress and fertility must be determined in futurestudies.

O-004

Whole Genome Amplification is efficient for CTG repeats length detection of PGD
for DM1
1,2

2

2

2

2

○Tomoyoshi Sakurai , Kou Sueoka , Kaori Takahashi , Suguru Sato , Kenji Sato ,Yasunori
2
Yoshimura
1

Saitama municipal Hospital Department of Obstetrics and Gynecology, Department of Obstetrics and Gynecology, Saiatama, Japan
Keio University School of Medicine, Tokyo, Japan

2

Introduction: Myotonic dystrophy type1 (DM1) is an autosomal dominant disorder, with an incidence of 1/8000. The geneis located on chromosome 19q13.3 and unstable expansion of the CTG repeats. This explains anticipation, that is increase in
severity PGD for DM1. PGD of DM1 has been performed by Nested PCR with linked polymorphic markers. However,carrying out PGD for DM1 is difficult for multiple reasons. First, the large expansion of CTG repeats is refractory to PCR
because of the high GC contents. Direct diagnosis exposes also risk of allele drop-out (ADO). The first single cell assay
developed for PGD for DM1 allowed selection of ―unaffected‖ embryos based on the presence in the biopsied cell of the‗normal CTG repeat‘ of the affected partner and on the ―normal CTG repeat‖ of the unaffected partner. Then it is necessarythat the healthy allele of the affected parent differs from the two short alleles (within 37 repeats) of the healthy parent.
Multiple Displacement Amplification (MDA) has been reported to yield large amounts of DNA from single cells. We
improved PGD for DM1 to using MDA for purpose of detect of two normal alleles. Material & methods: (Genetic test)
CTG repeats numbers of normal and affected alleles in both partners were analyzed before PGD. We analyzed the numberof CTG repeat of the client family using Gene Scan. Whole Genome Amplification(WGA) with MDA method and Gene
Scan were carried out for clients of PGD for DM1. MDA was performed using the GenomiPhiⓇ Kit on lymphocytes. Gene
Scan analysis was performed on the MDA product (5μl). This study aims to analyze the amplification rate and size of the
expanded CTG repeat number after MDA methods. Results: Single lymphocytes gave very high amplification efficiencies
for both protocols. Amplification rate by MDA was increased by this technique from 40% (4/10) to 60% (6/10). We
performed gene amplification by using MDA methods, the number of CTG repeat showed no change. Conclusion: A
repetition diagnosis of PGD for DM1 was enabled by using the MDA method which was non-PCR-based WGA. In
addition, we can expect possibility of a rise of the diagnosis efficiency and the reuse of the rejection embryo was shown by
using MDA. The MDA method showed the diagnosis efficiency that was higher than the Nested PCR method. WGA of
MDA was shown to be useful PGD for DM1.

240

●Oral Communications

O-005

Reducing multiple pregnancies by selection of a single chromosomally normal
blastocyst for transfer using array CGH (aCGH) analysis of 24 chromosomes
within 24 hours
1

2

1

2

○Zhihong Yang , Xiaohong Liu , Shala Salem , Jiaen Liu , Rifaat Salem
1

1

2

Pacific Reproductive Center, Torrance, California USA, Beijing Jiaen Hospital, 29 Zhichun lu, Beijing 10083, China

Introduction: Selection of a single viable blastocyst for transfer is the solution for reducing multiple pregnancies inthe stimulated IVF cycles. The objective of the present study was to evaluate the effect of selection of a single
euploidy blastocyst for transfer by using aCGH analysis of 24 chromosomes on clinical pregnancy outcomes.
Materials and methods: Two phases of studies were performed with an informed consent. In phase I, the efficacy ofaCGH screening of 24 chromosomes of trophectoderm cells derived from blastocyst biopsy within 24 hours wasevaluated. The trophectoderm cells were analyzed using the 24sure aCGH analysis of 24 chromosomes within 24
hours (BlueGnome, UK). The hatching rates were compared between the euploidy blastocysts and the anuploidy
blastocysts after blastocyst biopsy. In Phase II, the effect of transfer of a single euploidy blastocyst following
aCGH analysis on the clinical pregnancy outcomes was investigated. All patients were under 35 years old withnormal FSH levels on their first IVF cycles. After biopsy and aCGH analysis, a single euploidy blastocyst was
selected and transferred to each patient on day 6. A clinical pregnancy was defined as the presence of gestationalsac(s) and fetal heart beat(s) detected by ultrasound. IVF patients with similar demographic and clinicalcharacteristics were used as control. Significance of data was determined by Chi-square and t-tests. Results: 70.8%of the blastocysts were euploidy while 29.2% were aneuploidy. There was no significant difference in hatching
blastocyst rates between the euploid blastocysts and the aneuploid blastocysts (P>gt0.05). 91.2% (31/34) of thepatients had normal embryo(s) for transfer after the aCGH testing. 76.5% (26/34) of the patients became clinicallypregnant, 96.2% (25/26) of them was confirmed with ultrasound as singleton pregnancy while 3.8% (1/26) was
twin pregnancies.There was a significant difference in clinical pregnancy rates between the aCGH tested group andthe non-tested IVF control group (76.5% vs. 40.3%, P<lt0.05). Conclusion: Our data suggests that the aCHG
analysis of 24 chromosomes enables efficient differentiation of the euploidy blastocysts from the aneuploidy
blastocysts. Moreover, selection of a single chromosomally normal blastocyst for transfer by using the aCGH
screening of 24 chromosomes in fresh cycles may increase clinical pregnancy rates while reducing multiple
pregnancies.

O-006

Preimplantation genetic screening of eggs and blastocysts with CGH-microarray:clinical experience with the first 51 cases
1

1

1

1

1

○Stuart Lavery , Ben Lavender , Paul Knaggs , Anastasia Mania , Geoffrey Trew , Dagan Wells
1

2

2

IVF Hammersmith, Hammersmith Hospital, London UK, Reprogenetics Oxford UK

Introduction: The efficacy of preimplantation genetic screening has been one of the most keenly debated andcontroversial topics in assisted conception. Current evidence suggests that preimplantation screening of embryos at
the cleavage stage using multicolour FISH does not improve outcome. Recently the ESHRE PGS taskforce hasreported in a proof of principle study that screening of polar bodies using whole genome amplification andcomparative genomic hybridisation on micro-arrays is a reliable and robust method for determining thechromosomal status of the egg. Materials and methods: Polar body micro-array CGH is likely to be able to detect
where errors have a meiotic origin, but would not detect post zygotic errors leading to aneuploidy. Polar body
biopsy and micro-array analysis offers the advantages of more time to study the chromosomes, a more completekaryotype, and may be ethically more acceptable than embryo diagnosis. Biopsy of the blastocyst should be able todetect aneuploidies with maternal, paternal and post-zygotic origin. 53 patients with more than three failed cycles
each were eligible for our pilot study and all proceeded to egg collection. Results: 14 patients did not proceed togenetic analysis (less than three embryos). 7 patients had all aneuploid embryos and did not have a transfer. 4
patients had no CGH result but proceed to transfer of undiagnosed embryos, one conceived. 34 patients hadembryo transfer following euploid results and 10 have a clinical pregnancy (29% pregnancy rate/ET). Conclusions:True clinical validation of this technique requires a large multicentre prospective randomised clinical trial. Such a
trial is proposed by ESHRE with a completion date in 2013. Our initial pilot experience contributes somepromising data to this hotly contested area.

241

●Oral Communications

O-007

Most Motile Sperm Can Be Separated at the Bottom of the Microfluidic Channel ofa Plastic Microfluidic Sperm Sorter
1○Koji Matsuura , Keiji Naruse

2

1

Research Core for Interdisciplinary Sciences, Okayama University, Okayama, Japan, 2Cardiovascular Physiology, Graduate School of
Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan

Objective: A microfluidic sperm sorter (MFSS) is a simple chip device in which motile sperm are selected by two
gravity driven laminar flows within the microfluidic channel. Fluid mechanics simulation suggests that fluid
velocity is the highest at the center of the microfluidic channel and that it is difficult for sperm to swim across theinterface at the center. Because the observation of these phenomena using a conventional epifluorescencemicroscope is difficult, we employed a confocal microscope with a high-speed scanner. Here we investigated thenumber of sperm swimming across the interface in the 0.2 mm deep microfluidic channel of a plastic MFSS.
Materials and methods: Separation Procedures: Semen was left for at least 30 min to liquefy before processing andwas mixed with 2 μM Fluo-4 AM solution. Human Tubral Fluid (Irvine Scientific Inc., USA) was pipetted into thetwo outlets and one inlet of MFSS (Strex Inc., Japan), while the diluted semen was pipetted into the adjacent inlet.
Fluorescent Imaging: The intracellular concentration of Ca2+ was measured in stained sperm obtained from MFSS,
using a fluorescence microscope attached to a CSU10 confocal scanner (Yokogawa Electric Co., Tokyo, Japan).
The time resolution of each frame was approximately 50 ms. Twenty minutes after loading the semen sample, thenumber of sperm that swam across the field of view and the maximum fluid velocity during 1 min recording at thebottom, middle, and top of the channel were determined. Results: The number of motile sperm was highest at thebottom of the adjacent inlet. At the top and bottom of the MFSS channel, the fluid velocity had decreased and thesperm readily swam across the interface. These results suggest that the number of sorted motile sperm obtained
depends on the fluid velocity and the concentration of motile sperm at each level in the MFSS channel.
Conclusions: Motile sperm could be separated most efficiently at the bottom of the MFSS channel because thefluid velocity was sufficiently low to allow sperm to swim across the interface. Furthermore, the motile sperm wereconcentrated by gravity and sperm/geometry interactions. Human sperm concentrate at the bottom of the
microfluidic channel with decreased fluid velocity. This characteristic is an important consideration whenattempting to increase the number of sorted motile sperm.

O-008

Polymerization of Insulin-Like Growth Factor-Binding Protein-1 (IGFBP-1)
Potentiates IGF-I Actions in Placenta
1

2

2

2

2

○M Kabir-Salmani , H Shibuya , K Sakai , Y Wachi , M Iwashita
1

National Institute of Genetic Engineering and Biotechnology,Tehran, Iran, 2Department of Obstetrics and Gynecology, Kyorin
University School of Medicine, Tokyo, Japan

Background: Insulin-like growth factor (IGF)-binding protein-1 (IGFBP-1), the main secretory protein of deciduathat binds to IGFs and has been shown to inhibit or stimulate IGFs bioactivities. Polymerization, one of theposttranslational modifications of IGFBP-1, has been shown to lead to loss of inhibiting effect of IGFBP-1 on
IGF-I actions. The current studies were undertaken to elucidate the effects of steroid hormones on IGFBP-1
polymerization in trophoblast cell cultures. Materials and Methods: Placental tissues were obtained during legal,
elective procedures of termination of pregnancy performed between 7 and 10 weeks of gestation, and primarytrophoblast cells were separated. IGFBP-1 polymerization was analyzed by SDS PAGE and immunoblotting.
IGFBP-1 was polymerized when IGFBP-1 was added to trophoblast cell cultures. Polymerization of IGFBP-1 wasinhibited by the addition of anti-tissue transglutaminase antibody into the culture media. Results: There was anincrease in the intensity of polymerized IGFBP-1 bands with the addition of medroxyprogesterone acetate (MPA),
while no such difference was observed upon treatment with estradiol. MPA also increased the expression of tissuetransglutaminase on trophoblast cell membranes. IGF-I stimulated trophoblast cell migration, while IGFBP-1
inhibited this IGF-I-induced trophoblast response. Addition of MPA attenuated the inhibitory effects of IGFBP-1
on IGF-I-induced trophoblast cell migration. IGFBP-1 was polymerized by tissue transglutaminase on the cellsurface of trophoblasts, and MPA increased tissue transglutaminase expression on the cell surface and facilitated
IGFBP-1 polymerization. Conclusion: These results suggest that progesterone might facilitate polymerization of
decidua-secreted IGFBP-1 and increase IGF-I actions at feto-maternal interface, thereby stimulating trophoblast
invasion of maternal uterus.

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O-009

MicroRNAs associated with human embryo implantation defects
1

2

3○Ariel Revel , Hanna Achache , Juliet Stevens , Smith Yoav

4

1

Hadassah University hospital. Jerusalem, Israel, 2Institute for Drug Research, School of Pharmacy, Faculty of medicine, The Hebrew
University of Jerusalem, Jerusalem, Israel, 3University of Oxford, Oxford, United Kingdom, 4Genomic Data Analysis Unit, The Hebrew
University of Jerusalem, Jerusalem, Israel

Background: Repeated implantation failure (RIF) is a major problem encountered in in-vitro fertilization (IVF). We
have previously reported that RIF-IVF patients have a different endometrial gene expression profile during thewindow of implantation. Considering microRNA (miRNA) function in post-transcriptional regulation of gene
expression, the aim of the study was to confirm the involvement of miRNA in the defects of endometrial
receptivity. Methods: We used TaqMan miRNA array cards to identify the miRNAs differentially expressed in thesecretory endometrium of RIF-IVF patients as compared to fertile women and bioinformatics tools to identify theirpredicted targets and the molecular networks they may affect. Results: Comparing miRNA expression profiles, we
identified 13 miRNAs differentially expressed in RIF endometrial samples that putatively regulate the expressionof 3800 genes. We found that 10 miRNAs were overexpressed (including miR 145, 23b and 99a) and 3 were
underexpressed. Using our previous gene expression analysis, we paralleled miRNA 8211 mRNA expression
profiling. By this mean, we identified novel and previously characterized miRNA-regulated molecular pathways
such as adherens junctions, Wnt-signaling and cell cycle pathways. Consistent with the miRNA data, mRNA levelsof N-cadherin, H2AFX, netrin-4 and sFRP-4, belonging to the Wnt signaling and CAMs pathways were lower in
RIF-IVF patients. Conclusion: To our knowledge, this is the first study to evaluate the differential expression of
miRNAs in the secretory endometrium of RIF-IVF patients. We suggest that the RIF-associated miRNAs should be
exploited as new candidates for diagnosis and treatment of embryo implantation failures.

O-010

Pre-ovulation leptin serum as a marker for endometrial receptivity
11○Andon Hestiantoro , Marly Susanti , Atikah Barasila

2

1Division Reproductive Immuno-endocrinology, Department of Obstetrics and Gynecology, Faculty of Medicine University ofIndonesia, Dr. Cipto Mangunkusumo., 2Departmen of Hystology, Faculty of Medicine, University of Indonesia, Jakarta, Indonesia

Introduction: Having marker for predicting the endometrial receptivity would be the crucial point in calculating thetime for embryo transfer. Leptin is a peptide that play an important role in conducting and maintaining the goodcommunication between oocyte, embryo and endometrium. We are trying to find whether pre-ovulation leptin
serum would be the useful marker for predicting the endometrial receptivity.Material and Methods: This study was
conducted at Yasmin Reproductive and Fertility Clinic, Dr. Cipto Mangunkusumo Hospital, Jakarta, Indonesia.
Thirty-three infertile women were recruited for this study. Leptin serum levels were measured twice on day
11-13th, and on day 20-22th of menstrual cycle. Endometrial biopsy was taken in mid luteal phase for assessing
endometrial integrin αvβ3 immunohistochemically. Results: Positive correlation was observed between
pre-ovulation leptin serum and mid luteal phase leptin serum (r = 0.621). More over, pre ovulation leptin serum
showed a strong correlation with endometrial receptivity (p< 0.001). The best pattern of relationship between
endometrial receptivity and leptin level in this study is linear with the formula of endometrial receptivity score =1.813 x log (pre-ovulation leptin serum) -1.813. The leptin level that gave the best endometrial receptivity was
24.85ng/ml with 50.65 ng/ml as the maximal threshold. The multivariate analysis showed that body mass index,
endometrial thickness, and leptin serum level are the variables that have correlation to endometrial receptivity.
However, the only variable that showed significant corelation (p < 0.05) was pre-ovulation leptin serum (OR=1.08; CI 95%= 1.01-1.16). During this study, there were four patients who got pregnant with the integrin αvβ3
expression ranged between score 1.5 to 4.0. Conclusion: Pre-ovulation leptin serum can be widely used as a marker
to assess endometrial receptivity because it has a good predictive value. Moreover, it is easier to do and relatively
cheaper than integrin αvβ3 expression test.

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●Oral Communications

O-011

Application of Mechanical Stimuli Using a Microfluidic Air Actuation System:
Dynamic Embryo Culture
1

1○Koji Matsuura , Yuka Kuroda , Keiji Naruse

2

1

Research Core for Interdisciplinary Sciences, Okayama University, Okayama, Japan, 2Cardiovascular Physiology, Graduate School of
Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan

Objective: Mammalian embryos experience not only hormonal but also mechanical stimuli (MS), such as shear stress(SS), compression, and friction force, in the fallopian tube before nidation. Previous in vitro dynamic culture systems
have significantly improved embryo development compared to conventional static culture systems. However, a demeritof these systems is the need to use electric devices inside an incubator under humified conditions. In addition, we intend
to apply MS larger than those applied using these devices. To address these issues, we developed a dynamic embryo
culture system using air actuation and evaluated the applied MS and embryo culture results. Materials and methods: We
developed an air actuation system with microfluidic channels to apply MS by deforming a 0.1 mm thickpolydimethylsiloxiane membrane and evaluated SS applied to mouse embryos inside the microfluidic channel. Frozen 2cell stage embryos of the ICR mouse (Arc Resources Inc., Japan) were thawed and cultured in approximately 200 μl of
the potassium simplex optimized embryo culture medium for 3 days at 37 C under a humidified atmosphere of 5% CO2.
The number of cultured embryos was from 10 to 13. Results: When syringe velocity (Vs) was 0.5 mm over seconds, theobserved embryos rotated and did not slide. When Vs and fluid velocity increased, the embryos slipped, did not come incontact with the floor, and were not compressed because of the fast translation. We conclude that different types andamounts of MS can be applied to the embryos by changing Vs. Maximum SS applied using this system wasapproximately 10 times higher than that using previous dynamic culture systems such as the tilting embryo culture
system and microfunnel. We compared embryo development from the 2 cell stage to blastocyst stage between the staticand dynamic cultures in the medium channel. Dynamic cultures significantly improved the blastocyst development rate[dynamic 74% (n = 126); static 62% (n = 118); P < 0.05]. The average number of cells in the blastocysts cultured underdynamic conditions was 83 cells (n = 54, ± 3), while that under static conditions was 76 cells (n = 51, ± 3). A significant
difference was observed in the average cell number between the two groups (P < 0.05). Conclusions: We demonstrated
that this device could be used for dynamic embryo culture in a microfluidic channel. MSs applied using this system weresimilar to those generated in the physiological environment of the oviduct.

O-012

Caffeine treatment influences in vivo and in vitro fertilization, embryo development
and morphology in mice
1

1

1

1

○Yoshimasa Yokota , Mikako Yokota , Hidemi Yokota , Setsuko Sato , Yasuhisa Araki
1

2

2

Yokota Maternity Hospital, The Institute for Advanced Reproductive Medical Technology, Gunma, Japan

Introduction: We previously reported that pregnancy rates and testosterone levels in the follicle were significantly lower inpatients that habitually drink coffee. Caffeine thus appears to affect regulation mechanisms related to testosterone releaseand oocytogenesis. Therefore, in the present study, we evaluated effect to produce oocytes by caffeine in vivo and embryo
growth features after in vitro fertilization (IVF) using oocytes after long-term caffeine treatment in mice, and we evaluatedthe direct influence on embryo growth using culture media containing different levels of caffeine.Material and Methods:
Mature, female ICR mice were divided into three groups and were treated with intraperitoneal injection of caffeine for one
month at 0, 0.1 or 1 mg/day equivalent. Oocytes were collected from oviducts at 14 h after superovulation with 10IU
PMSG and 5IU hCG, and used these for IVF, and then observed embryo development. In an independent experiment, after
IVF, we evaluated the development of embryos to blastocyst formation and morphology using media that directly included
different caffeine concentrations at 0, 0.05 and 1.0 mg/ml. Results: Number of oocytes dose-dependently decreased after
caffeine treatment. Mean oocytes per mouse in the 1 mg/day group was significantly lower (17.7) than in the 0 mg/day
control group (39.0) (P<0.05).In vitro fertilization rate did not significantly differ from 0 to 1.0 mg/ml; however, theformation of blastocysts at 120 h after insemination was significantly lower with higher caffeine levels; 49.8% in the 0.1
mg/day group and 38.2% in the 1.0 mg/day group (vs. 64.0% in controls) (P<0.05). When caffeine was directly added to
culture media, blastocyst formation rate was significantly lower (0%; 0/81) at 1.0 mg/ml caffeine, as compared with 98.8%(80/81) in the 0.05 mg/ml group (P<0.05) and 97.4% (75/77) in the control group (P<0.05). Conclusions: Caffeine
treatment prior to retrieving oocyte influences oocytogenesis, and reduces the number of oocytes and blastocyst formation
rate using picked up oocytes in vitro. The results also show that caffeine in media has direct negative effects on embryo
development to blastocyst formation in vitro. Thus, it suggests that caffeine affects embryo development as well as
producing oocytes in the body. Further studies are needed in order to understand the mechanisms responsible for theseeffects.

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●Oral Communications

O-013

Dynamic analysis of compaction initiation in human embryos using time-lapse
cinematography○Kyoko Iwata, Keitaro Yumoto, Akifumi Imajo, Yumiko Iba, Yasuyuki Mio
Mio Fertility Clinic, Yonago, Japan

Objective: Compaction is the change in cellular adherence that indicates the first morphological event in embryonicdifferentiation. Although it was reported that it occurs at the late eight-cell stage in mouse embryos, it is still unknown inhuman. We used time-lapse cinematography to examine the timing of initiation of this event in cultured human embryos.
This study analyzed the etiology as well as the fate of those embryos showing early initiation of compaction in humanembryos obtained from TLC. Materials and Methods: We developed a new system for time-lapse cinematography (TLC),
which uses an inverted microscope with differential interference contrast (DIC) optics and a micromanipulator covered witha handcrafted chamber of acrylic resin. The temperature was maintained at 37±0.2°C and pH at 7.37±0.05 by controlling
CO2 flow. Digital images of the cultured embryos were acquired for 5 days with an exposure time of 1/20 second. Of 119
donated embryos frozen at an early developmental stage (pronuclear stage to 4-cell stage), 109 successfully developed tothe blastocyst stage and could be analyzed. Result: Our TLC analysis revealed that compaction occurred in 115 embryos.
Initiation of compaction occurred from the 4- to 16-cell stage, with the 8-cell stage the most common (26/115; 22.6%). Of
those embryos, 99 (86.1％) embryos initiated compaction after the 8-cell stage and 49 (49.5％) developed into good-quality
blastocysts. On the other hand, of the 16 (13.9％) embryos in which compaction was initiated before the 8-cell stage, 15(93.8%) presented multinucleated blastomeres, and only 3 (18.8%) developed into good-quality blastocysts. The remaining
13 (81.3%) developed into poor-quality embryos. There was a significant difference in the rate of development into
good-quality embryos between embryos that initiated compaction after the 8-cell stage and before the 8-cell stage (P<0.05).
Ten of the embryos that initiated compaction before the 8-cell stage (62.5％) showed interruption of blastomere cleavage
after forming of the cleavage furrow, followed by multinucleation in the blastomere. There was no difference in timerequired from embryo thawing to the initiation of compaction between the embryos where compaction started before the8-cell stage and those that initiated compaction after the 8-cell stage. Conclusions: Our study suggested that the initiation ofcompaction is more likely to occur in human embryos at the 8-cell stage or later and that early initiation of compaction isstrongly associated with poor quality in human embryonic development. Because most of the multinucleated blastomeres
observed here failed to complete normal cytokinesis, early initiation of compaction might occur due to aberrant cytokinesis
followed by karyokinesis.

O-014

Chromosomal analysis of cleavage embryos and blastocysts derived fromtripronuclear zygotes after ICSI.
○Shimpei Mizuta, Nobuhiko Kataoka, Hiromi Hashimoto, Yasushi Kuroda, Shoji Kokeguchi,
Masahide Shiotani
Hanabusa Women's Clinic, Hyogo, Japan

Introduction: In IVF and ICSI, normal fertilization is determined by the presence of two distinct pronuclei (2PN) at 16-20
hours after insemination (Day-1). Occasionally, three pronuclear zygotes (3PN) are observed. 3PN after IVF, in most cases,
is treated dispermic fertilization with the chromosomal abnormality, triploid. It has been reported that 70% of ICSI 3PN
cleavage embryos were polyploid. However some of them have been showed diploid. If these embryos can be estimateddiploid morphologically, they might be useful for embryo transfer for the patient that does not have a lot of oocytes orembryos. In this study, we analyzed the chromosomes of ICSI 3PN cleavage embryos and blastocysts. Materials andMethods: In cleavage embryos, among 4016 oocytes after ICSI from July 2008 through May 2009, we conductedchromosomal analyses of Day-3 37 embryos defined as 3PN at Day-1after ICSI. In blastocysts, 112 3PN of 5171 oocytes
were observed at Day-1after ICSI from July 2008 through May 2010. 102 of 112 embryos were cultured to Day-5 or 6, andthen 23 blastocysts were examined for chromosome. The chromosomes of cleavage embryos and blastocysts were
examined by fluorescence in situ hybridization technique with 18, X and Y probes. Results: One-hundred-twelve of 5171
oocytes (2.2%) after ICSI were found 3PN. The percentage of BLs formed from 3PN was 24.5% (25/102). The
chromosome analysis showed that, 8.1% (3/37), 29.7% (11/37), 32.4% (12/37), 2.7% (1/37), 21.6% (8/37) and 2.7% (1/37)
of the ICSI 3PN cleavage embryos were 2n, 3n, mosaic included 3n, mosaic included 4n, n, chaotic mosaic and otherabnormality, respectively. The polyploidy embryo rate was 67.6% (25/37). Y chromosome was detected in 45.9% (17/37)
of the ICSI 3PN cleavage embryos. 4.3% (1/23), 52.2% (12/23), 26.1% (6/23), 13.0% (3/23), and 4.3% (1/23) of the ICSI
3PN blastocysts were 2n, 3n, mosaic included 3n, chaotic mosaic, and other abnormality, respectively. The polyploidy
blastocyst rate was 78.3% (18/23). Y chromosome was detected in 56.5% (13/23) of the ICSI 3PN blastocysts.
Conclusions: In the ICSI 3PN cleavage embryos, the prevalence of 2n was 8.1% and the chromosomal abnormality was
91.9%. In the ICSI 3PN blastocysts, 2n was 4.3% and the chromosomal abnormality was 95.7%. ICSI 3PN was confirmed
to have a high chromosomal abnormality rate in both of cleavage embryos and the blastocysts. Therefore, even if ICSI 3PN
zygote develops to the blastocyst, it should not be useful for embryo transfer.

245

●Oral Communications

O-015

THE DYNAMIC PROCESS OF SPERM PENETRATION OF THE HUMANOOCYTE ANALYZED USING TIME-LAPSE CINEMATOGRAPHY○Yoshiteru Kai, Kyoko Iwata, Keitaro Yumoto, Akifumi Imajo, Yumiko Iba, Yasuyuki Mio
Mio Fertility Clinic, Yonago, Japan

Objective: Using time-lapse cinematography (TLC), we analyzed the dynamic process of human sperm penetration
to investigate the mechanisms blocking polyspermy after sperm attachment. Materials and Methods: Between July
2004 and April 2008, TLC was performed on single, randomly selected oocytes after insemination (Mio, 2008).
The study focused on oocytes where the movements of the leading sperm (the one that penetrated the zona
pellucida (ZP) most deeply) and the following sperm were captured in the same field. Sperm movements were thenanalyzed from the time of entry into the ZP to the incorporation of the leading sperm. Results: We observed theprocess of sperm penetration and the sperm entry point in 22 embryos. In 3 of these embryos (target embryos), wewere able to capture the movements of the leading and following sperm. Sperm attached to the oocyte membrane
approx 96 minutes after insemination and 37 minutes later, the sperm head decondensed and disappeared. There
was no difference in the time-course of the fertilization process of the target embryos and the other embryos
observed. In the 3 target embryos, both leading and following sperm proceeded forward until the leading sperm
attached on the ooplasmic membrane. The following sperm penetrated 12.9μm of the ZP on average and, despite
their active tail movements, stopped their progression within 10 seconds of the leading sperm attaching on themembrane. Conclusions: To date, the primary block to polyspermy in the mammalian oocyte is described as a zona
reaction induced approximately 5 to 8 minutes after oocyte activation by sperm incorporation. In this study, 10
seconds after sperm-oocyte attachment, the following sperm were unable to continue further despite their sustained
tail motility. This suggests that an unknown process has prevented the progression of the following sperm. Further
investigations are now on going.

O-016

Depletion of Plk1 delays cell cycle progression in endometrial cells derived fromendometriosis
1,2

3

4

2

2

Li Tang , Tian-Hua Zhou , Jian-Zhong Sheng , Yan-Ting Wu , Ming-Yue Dong , ○He-Feng
2
Huang
1

The First People's Hospital of Yunnan Province, 2Women's Hospital, Zhejiang University, School of Medicine,
Zhejiang University, School of Medicine, 4University of Calgary, Canada

3

The ectopic (Ect) and eutopic (Eut) endometrial cells (ECs) derived from Endometriosis (EMS) displayed similarcellular biological behaviors to malignant cells, but their specific cellular pathogenesis remains to be elucidated. To
investigate the pathological mechanism of endometriotic cells, firstly, we corroborated a higher polo-like kinase 1(Plk1) expression in ECs from women with EMS, either in Ect or Eut emdometrium. Furthermore, the roles ofendogenous Plk1 in ECs growth and proliferative regulation were investigated. Three kinds of ECs (Ect and Eut
ECs from women with EMS and Eut ECs from normal women) were stably cultured and transfected with small
RNA interference (siRNA) targeting Plk1. We observed retarded growth rate and reduced proliferation in depletionof Plk1 ECs as compared with control mock-transfected ECs. Cell cycle analysis displayed that ECs expressing
Plk1-specific siRNA accumulated during G2/M phase of the cell cycle. Further analysis demonstrated that,compared with normal Plk1-depleted ECs, endometriotic Plk1-depleted ECs showed a significantly delayed
induction of cyclin B1. These results suggest that knockdown of Plk1 in ECs from EMS may disturb cellproliferation and delay cell cycle progression by inhibiting cyclin B1 expression.

246

●Oral Communications

O-017

Male fetal DNA detection in maternal serum from pregnant cynomolgus monkeys(Macaca fascicularis) in an established breeding colony
1

2

3

3

1

○Lubna YASMIN , Jun-ichiro TAKANO , Yasushi NAGAI , Junko OTSUKI , Tadashi SANKAI
1

2

Tsukuba Primate Research Center, National Institute of Biomedical Innovation , Department of Research Resource Developments,
The Corporation for Production and Research of Laboratory Primates, 3Nagai Clinic

Introduction: Determination of fetal sex at an early stage of gestation is essential for the prenatal diagnosis of certain
inherited dominant genetic diseases such as the male-specific disorders Duchenne muscular dystrophy and aneuploidy.
As human and nonhuman primates are similar in their development, monkeys are often used as a model for fetal
development in biomedical research. There has been no information about the presence of fetal (f) DNA in maternal
serum during gestation for the most widely used model in biomedical research, the cynomolgus monkey. Breeding of thecynomolgus monkey is not controlled seasonally as it is in the rhesus monkey. The aim of this study was to develop an
efficient method for detecting fDNA in monkey serum at an early stage of gestation to diagnose gender-specific genetic
disorders. Material & Methods: We used quantitative real-time PCR with the TaqMan system to analyze maternal serum
obtained from pregnant cynomolgus monkeys in an established monkey breeding colony. Primers and probes for the
cynomolgus monkey-specific SRY gene and the Y-chromosome-specific multicopy marker sequences DYS14 were
designed by using Beacon designer 7.9 software. The presence of SRY and DYS14 was determined in five cynomolgus
monkeys at the 5th, 12th and 22nd weeks of gestation. All the animals were examined by ultrasonography, and sex was
confirmed at delivery. Results: A real-time quantitative generic multiplex PCR assay was developed that is capable ofdetecting the specific region of SRY and DYS14, making it possible to quantify the number of gene copies in each
multiplex reaction. We report the 100% specificity and 100% sensitivity of the probe for detecting the Y-chromosome
sex-determining region at the 12th and 22nd gestational weeks in the monkeys studied. At the 5th gestational week, weobserved <100% sensitivity for detecting male fDNA. Conclusions: We present a specific, highly sensitive method for
determining and quantifying male specific fDNA in pregnant cynomolgus monkeys. The lower sensitivity at the 5th
week of gestation might be due to a lesser amount of fDNA during early pregnancy.

O-018

Clinical outcome of 852 tubal factor infertility patients without ART after
Falloposcopic tuboplasty.
○Shoji Kokeguchi, Nobuhiko Kataoka, Seiji Ogata, Satoshi Yamada, Yukiko Matsumoto,
Masahide Shiotani
Hanabusa Women's Clinic, Hyogo, Japan

Introduction: Falloposcopic tuboplasty (FT) has been recognized as a highly useful instrument of both an assessment of
tubal luminal lesion and a surgical technique of recanalization of tubal occlusion and stenosis. We have used FT as adiagnosis and a treatment for about 1500 over tubal infertility patients since 2003 at our clinic. The aim of this study was toevaluate the pregnancy outcome without ART after FT and the effectiveness and safety of FT. Materials and Methods. FT
was performed for the 1468 patients with bilateral or unilateral occlusion and stenosis from January 2005 throughDecember 2010. These patients had been diagnosed by hysterosalpingography, hysteroscope and hydrotubiation. All FT
was performed with the procedure of transcervical balloon tuboplasty by intaravenous anesthesia in outpatient base. Lesions
in the tubal lumen were observed falloposcopically during retrograde imaging after complete recannulation. 852 patientswere able to follow up, and 253 of them have been pregnant by natural intercourse or artificial insemination. Patients withhydrosalpings, severe male factor and ART followed FT were excluded from this study. Results: Mean age of the patientswas 33.5 years old (ranged from 22 to 49 years old) Clinical pregnancies occurred in 253/852 patients (29%). All
pregnancies were singletons and the miscarriage was 7.5%. The pregnancy rate decreased with the age. The highest, 50%(3/6), pregnancy rate was seen for the 25 year old cases. Under 38 years old, the pregnancy rate was over 22 %. The oldest
case of conception was 41 years old, but she had a miscarriage. The mean pregnancy period after FT was 3.5 months. The
pregnancy cases occurred within the first month after FT. Within 6 months after FT 85.7% (216/252) of the women became
pregnant. Ectopic pregnancy was 1.6% (4/253). The 250 out of 599 cases that had not conceived were conducted to ART
and at present about 82.4% have become pregnant. Conclusion: Although ART has been used for women with tubal lesion
as a standard treatment, we apply FT as an initial treatment before ART. Our results show that this FT technique is usefulfor the patients with tubal infertility and does not cause an increase of ectopic pregnancy. FT might be recommended to theselected patients with tubal infertility under 40 years old before conducting IVF. And it is also recommended to reconsider
the therapeutic plan for these patients about 6 months over after FT.

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●Oral Communications

O-019

Outcomes of vitrified-warmed embryo transfer (FET) cycles: single blastocyst
transfer versus double blastocyst transfer○Elena S. Miadova
Perinatal Medical Center, Moscow, Russia

Introduction: Vitrification, an ultra-rapid freezing technique, is an optimal cryopreservation procedure for embryos.
Since 2008 while we've been using the Cryotop method for human embryo cryopreservation we achieved highclinical pregnancy rate after transferring two vitrified-warmed blastocysts (double FET). But these optimistic
results came across with the high rate of twin pregnancies that has a negative impact on the perinatal outcome of
the IVF programs. In order to reduce the multiple pregnancies rate we began to perform single vitrified-warmed
embryo transfer (single FET). The purpose of the present retrospective study was to compare the clinical outcomesof single versus double FET cycles. Materials and methods: 237 patients were included in the study. All goodquality supernumerary day 5-6 blastocysts were vitrified by the Cryotop method and then warmed for carrying out
embryo transfer. We analyzed 269 FET cycles performed from January 2010 to January 2011. Assisted hatching
was used to remove ¼ part of zona pellucida after blastocyst warming. No significant differences were observed
between the survival rate in single and double FET (100% and 98.8%). Endometrium for embryo implantation was
prepared by the hormonal replacement therapy or in natural cycle. The main outcome measures were clinicalpregnancy rate, ongoing pregnancy rate and implantation rate. Results: 225 and 44 ET were carried out in double
and single FET cycles respectively. The differences in the endometrium method preparation between groups were
not significant. The clinical pregnancy and ongoing pregnancy rates were higher in double versus single FET(62.7% vs 52.3% and 53.3% vs 45.5%, respectively). Implantation rate was 52.3% in single FET cycles and 41.1%
in double FET cycles. The multiple pregnancy rate was significantly lower in single compared to double FETgroup (4.3%: 1 case of monozygotic twins vs 31.2%: 43 dizygotic and 1 monozygotic twins, respectively).
Conclusion: The Cryotop method keeps high implantation potential of blastocyst. The transfer of one
vitrified-warmed blastocyst gives us an opportunity to achieve high pregnancy rates while reducing the multiple
pregnancy rate making IVF-procedures rather more patient friendly.

O-020

Are Mild ART-derived blastocysts more favorable than conventional COH-derived
ones?
1

1

1

1

2

○Yasushi Takai , Ken Ohara , Shigetaka Matsunaga , Masahiro Saito , Osamu Ishihara ,1
Hiroyuki Seki
1

Saitama Medical Center/Saitama Medical University, Saitama, Japan, 2Saitama Medical University, Saitama, Japan

Introduction: To evaluate the efficacy of assisted reproduction with mild stimulation using clomiphene citrate(CC-ART), we compared the outcome of frozen-thawed transfer of blastocysts obtained by CC-ART with those by
conventional controlled ovarian hyperstimulation protocol (COH-ART). Materials and methods: CC-ART was
conducted exclusively for patients with COH-ART failure or poor ovarian reserve. In our CC-ART protocol, daily
50 mg clomiphene citrate was started on the 3rd day of menstrual cycle and a couple of low dose gonadotropin
were administered after the 8th day. Blastocysts with Gardner's grade 3BB or better were cryopreserved without
fresh embryo transfer, thawed and transferred in natural cycle or HRT regimen. Results: In 532 CC-ART cycles,
blastocysts were cryopreserved in 184 cycles (34.6%). Pregnancy rate (PR) of single frozen-thawed blastocyst
transfer (SBT) after CC-ART (27.8%; 44 in 158 cycles) was significantly lower than those after COH-ART(35.4%; 336 in 950 cycles), although patients who had SBT after CC-ART (38.6±3.3 yo) were significantly older
than those after COH-ART (35.1±3.8 yo). When PR was compared in age-matched condition, however, nosignificant diffirence was detected (45.5% vs 38.3% in 30-34 yo, 28.8% vs 33.1% in 35-39 yo, 21.1% vs 31.7% in
40- yo, for CC-ART vs COH-ART, respectively). Conslusions: PR of blastocysts obtained by CC-ART wascomparable with those by COH-ART, even when CC-ART was limited for patients with COH-ART failure or poor
ovarian reserve. Age- and AMH-matched RCT may remain to be conducted.

248

●Oral Communications

O-021

Progress towards a universal warming method after vitrification
1

1,2

1

1

○Aniko Reichart , Gabriella Uhereczky , Miklos Sipos , Vince Forgacs , Gabor Vajta

21

Forgacs Institute, Budapest, Hungary, 2BGI Shenzhen, Yatian District, Shenzhen, China

Introduction: Establishment of various vitrification techniques with diverse carrier tools and solutions may create serious
problems when cryopreserved samples are transported from one clinic to the other, and routinely used vitrification techniques
differ between the two institutions. The purpose of our work was to test the possibility to establish a universal warming methodthat can be used after vitrification performed with various techniques. Material and Methods. In Experiment 1, D5 blastocysts
and MII phase oocytes were exposed to in-house media, 7.5-7.5% ethylene glycol (EG) and dimethylsulphoxide (DMSO) for
15 min, then 16-16% EG-DMSO and 0.5 M sucrose for 1 min before loading into open pulled straws (OPS) and cooling.
Warming was performed in 1M sucrose for 1 min, then in 0.5 M sucrose for 3 and in holding medium for 2x5 min,respectively. Samples were transferred in large amount (20 µl) medium from the concentrated to the diluted solution.In
Experiment 2, D3 embryos and blastocysts vitrified with the MediCult Vitrification kit and parameters were warmed either byusing the original MediCult solutions and parameters, or in-house media and parameters as described above. The age ofpatients did not differ significantly between the two groups. Results. In Experiment 1, the re-expansion rate of blastocysts was
100% (7/7). Survival and developmental rates to morula/blastocysts stage after vitrification of oocytes were 100% (9/9) and
30% (3/9). In Experiment 2. survival of morulae and re-expansion rates of blastocyst warmed in Medicult versus in-house
media were 92% (55/60) and 66% (45/68) versus 98% (47/50) and 64% (27/42) respectively Pregnancy rates achieved withtransfers of embryos warmed in Medicult versus in-house media were 21% (12/41) versus 37% (11/30), respectively.
Discussion Results of Experiment 1 have demonstrated that in-house media could be successfully applied for vitrification andwarming of both MII oocytes and blastocysts. In Experiment 2, warming with in-house media resulted identical survival rates
than those achieved with the original Medicult warming media, and pregnancy rates tended to be higher. As the Medicult
vitrification kit uses different cryoprotectants (propylene glycol instead of DMSO) and equilibration parameters different from
our in-house media, these results indicate that establishment of an universal warming method is a realistic perspective andresearch should be conducted towards this direction.

O-022

Failure of pronucleus formation in in vitro matured and fertilized oocytes from
SOD1-deficient mice
1

1

1

2

○Naoko Kimura , Yasuko Sato , Manami Suenaga , Satoshi Tsunoda , Junichi Fujii

2

1

Laboratory of Animal Reproduction, Graduate School of Agricultural Sciences, Yamagata University, Tsuruoka, Japan, 2Departmentof Biochemistry and Molecular Biology, Graduate School of Medical Science, Yamagata University, Yamagata, Japan

Introduction: Higher levels of intracellular reactive oxygen species (ROS) are detected in cleavage-stage embryos producedin vitro, compared with in vivo, and are related to early developmental arrest or cell death. We have previously shown total
two-cell arrest of in vitro-fertilized embryos from superoxide dismutase 1-deficient mouse (SOD1KO) oocytes without
mitochondrial malfunction under culture with 20% O2, indicating that the mechanism of cell cycle regulation might be atarget of elevated ROS. This study was conducted to investigate effects of a SOD1 deficiency during IVM on fertilizationand the following development. Material and Methods: Immature cumulus-oocyte complexes (COCs) were retrieved fromICR SOD1KO and wild-type mice (WT) after eCG administration. The COCs underwent IVM for 18 h under several
culture conditions, followed by IVF with WT sperms. In parallel, matured oocytes were treated with 10 mM SrCl2 forparthenogenetic activation (PA). The oocytes were assessed the nucleus stage at 6 h after insemination or PA. To observe
the early events of egg activation, cortical granules (CGs) were detected by lectin staining. Expressions of CaMK II werealso detected by immnofluorecence staining. Results: Under culture with 20% O2, SOD1KO oocytes were completely
prevented from attaining to two-cell stage. Most of the SOD1KO oocytes were arrested at Met II to Telo II at 6 h after
insemination regardless of the high frequency of sperm penetration. Culture with antioxidants or hypoxia slightly improved
cleavage. PA did not remove impairment of pronucleous formation in SOD1KO oocytes, indicating that cascades throughsperm factors were not related to this impairment. The CGs distributed on the peri-membrane of the Met II oocytes, apartfrom over-the-spindle, were slightly weakened after egg activation. Phospho-CaMK IIαβ was localized in the cytoplasm
with a strong presence on the Met plate. CaMK IIγ was detected on the peri-membrane of the Met II oocytes, apart fromover-the-spindle, and was present spottily in the cytoplasm after egg activation. The localizations of CGs, phospho-CaMKIIαβ and CaMK IIγ were similar between SOD1KO and WT oocytes. Conclusions: The oxidative stress caused by a SOD1
deficiency during oocyte maturation causes the failure of pronucleus formation after fertilization. This impairment seems tobe irrelevant to the abnormality of egg activation events, implying an injury to the cell cycle regulation of the meiosis II exit.

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●Oral Communications

O-023

A randomized double-blind comparative study between two different laser assisted
hatching in the frozen-thawed embryo transfer at the cleavage stage○Lei Ao
Yunnan First People Hospital, Kunming, China

BACKGROUND: Laser Assisted Hatching (LAH) in the frozen-thawed embryo transfer (FET) cycles increases theimplantation and pregnancy rates. However, Little information exists in the literature regarding the efficacy ofdifferent LAH. The object of this study was to evaluate the efficacy of two different LAH in the frozen-thawed
embryo transfer at the cleavage stage. METHODS: A total of 554 FET cycles using LAH was undertaken in ourcentre between October 2009 and June 2010. Embryos were cryopreserved at the cleavage stage. On the day of
FET, 554 cycles were randomized according to a computer-generated randomization list into the zona drillinggroup and zona thinning group. The patients and the clinicians were blinded to the group assigned. In the zona
drilling group, the drilled hole was 1.5 times the thickness of zona pellucida; In the zona thinning group, the outerhalf of the zona pellucida over a quarter of the diameter of zona was removed using a non-contact laser.
RESULTS: In the zona drilling group, the clinical pregnancy rate of 39.4% with an implantation rate of 22.4%. In
the zona thinning group, the clinical pregnancy rate of 42.1% with an implantation rate of 24.9%. All data of
pregnancy rate and implantation rate in the zona thinning group were slightly higher in comparison with the zona
drilling group, although there was a non-significant difference of pregnancy rate and implantation rate between thezona drilling group and the zona thinning group. CONCLUSIONS: In order to avoid the damage of the embryos in
the LAH, the zona thinning should be performed routinely to replace the zona drilling in the FET cycles at thecleavage stage.

O-024

A trial for observation of chromosome dynamics in human embryos using alive-cell imaging system
1

1

1

1

1

○Yoshiharu Nakaoka , Shu Hashimoto , Ami Amo , Keijiro Ito , Yoshiharu Morimoto ,2
Kazuo Yamagata
1

IVF Namba Clinic, Osaka, Japan, 2Research Institute for Microbial Diseases, Osaka University(Introduction) Chromosome analysis is generally performed using fixed samples. Recently, the development of aconfocal imaging system that includes an embryo culture system has made it possible to obtain time-lapse imagesof chromosome dynamics in mouse (Yamagata et al. 2010). In this study, we attempted to observe chromosome
dynamics and cytokinesis in human embryos using an all-in-one confocal imaging system. (Material & Methods)
Eighteen pronuclear embryos intended for disposal were used after obtaining the informed consent of the patientsand the approvals of the IVF JAPAN and Japan Society of Obstetrics and Gynecology research ethics committees.
A mixture of mRNAs encoding enhanced green fluorescent protein coupled with α-tubulin and monomeric red
fluorescent protein I fused with histone H2B was injected into the cytoplasm of the pronuclear embryos using a
Piezo drive manipulator. The pronuclear embryos were cultured under the atmosphere of 5% O2, 5% CO2 and 90%
N2. Time-lapse images were captured at 15-minute intervals until the day 3 stage using an all-in-one confocal
imaging system (CV1000, Yokogawa Electric Corp.). (Results) Fluorescent signals from tubulin and histone weredetected in all RNA-injected embryos around first mitosis. However, signals at first mitosis of some embryos were
not so clear because the fluorescent protein had not yet been translated from mRNA. (Conclusions) We succeededin observing chromosome dynamics using an all-in-one confocal imaging system in human live embryos. The
observation of chromosome dynamics in human embryos using a live-cell imaging system may help to elucidate
the mechanisms and incidence of chromosome aberrations such as aneuploidy and polyploidy.

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O-025

Autologous transplantation of very thin ovarian fragments which preserve theovary's main cortex structure lead to successful pregnancy
1

1

1

1

1

2

○Ariel Revel , Meital Lebovich , Alex Simon , Neri Laufer , Einat Eizenmann , Eduardo Mitrani
1

Hadassah University hospital. Jerusalem, Israel, 2The Alexander Silberman Institute of Life Sciences, hebrew university, Jerusalem,
IsraelIntroduction: Ovarian cortex was cryopreserved in a young thalassemic patient prior to premature ovarian failure
associated with stem cell transplantation. Aim: to restore fertility using frozen thawed ovarian cortex. Materials:
Eleven in vitro fertilization (IVF) cycles were then performed following conventional ovarian-strip transplantation;
however, the ovarian response was poor. In parallel, we explored, in a rat model, the use of micro-organs (MOs),
i.e. microscopic ovarian fragments, which preserve ovarian cortex architecture allowing for sufficient diffusion ofgases and nutrients. Results: We found that rodent ovarian MOs induced angiogenesis and remained viable after
autologous transplantation. Subsequently, we reimplanted MOs prepared from autologous thawed ovarian strips
into our patient, resulting in a successful delivery. Conclusions: Precision cutting is important in order to preserve
ovarian cortex architecture. The facts that the resulting fragments are of uniform thickness and only a few hundred
microns thick, ensure reproducibility and appropriate and homogeneous diffusion of gases and nutrients to all cellcomponents. Furthermore, very thin (350μ) ovarian fragments secrete a whole range of angiogenic factors which
ensure that a vascular network is rapidly formed.

O-026

Important implications of advanced glycation end-products (AGE) in poor ART
outcomes and a novel successful therapy for very severe ART patients withsitagliptin possibly by decreasing AGE.
1

2

1

1

1

○Masao Jinno , Masayoshi Takeuchi , Aiko Watanabe , Jun Hirohama , Naohisa Hatakeyama ,1
Rie Hiura
1

Women's Clinic Jinno, Tokyo, Japan, 2Kanazawa Medical College, Ishikawa, Japan

Objective: Advanced glycation end-products (AGEs) are accumulated with ageing, diabetes mellitus and polycystic
ovary syndrome, playing a pathogenic role. We showed adverse effects of AGE accumulation on ART outcomes instudy I and in study II we improved ART outcomes in very severe ART patients by decreasing postprandial
glycemic levels and AGEs with sitagliptin, a new antihyperglycemic agent. Materials and Methods: Study I: Toxic
AGE (TAGE), pentosidine (Pent), and carboxymethyl lysine (CML) in blood and follicular fluid (FF) were
measured in 157 ART-patients. We analyzed associations of AGE with ART outcomes. Study II: Sitagliptin, 50
mg/day, was administered for 2 months before ART in 41 very severe ART repeaters with 7.6±0.7 (SEM) attempts
of previous failed ARTs and 41.0±0.5 years of age. 75g-oGTT was normal and borderline in 39 and 2 patients,
respectively. All patients had failed with metformin. TAGE measurements and oGTT were done before and one
month after sitagliptin in 36 patients. Results: Study I: TAGE, Pent, and CML in FF, and TAGE in serum showedsignificant negative correlations with follicular and embryonic develoment and fertilization. Among 12 factors,
only age, Pent in FF, and TAGE in serum were correlated with ongoing pregnancy significantly. Women withserum TAGE above 7.24 U/mL showed decreased oocyte numbers and ongoing pregnancy rates, even withyounger age or lower day-3 FSH. Study II: Sitagliptin significantly decreased plasma glucose levels at 30, 60, and
120 minutes after 75 g glucose in oGTT, but not fasting levels. Serum TAGE was decreased, unchanged orincreased in 17, 6 or 13 patients, respectively (groups D, U or I). The number of day-2-superior-embryos wasincreased by sitagliptin in 47 and 50% of patients in groups D and U, respectively, being significantly higher than
8% in group I. Consequently, 3 ongoing pregnancies and 2 abortions were achieved in 41 ART with sitagliptin(rates of clinical and ongoing pregnancy: 12% and 7.3%, respectively), while 8 abortions alone in their 244
previous ART attempts without sitagliptin (3.2% and 0%, respectively). Conclusions: Serum TAGE and FF Pentaccumulations correlated highly with poor follicular and embryonic developments and with unlikelihood of
ongoing pregnancy. Serum TAGE predicts poor ART outcomes independently of age and day-3 FSH. Decreasing
postprandial glycemic levels and TAGE by sitagliptin is possibly a novel therapy for ovarian dysfunction.

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●Oral Communications

O-027

A novel long protocol of GnRH agonist and hMG regimen: a dramatical increase inpregnancy rate by induction of diminished but significant mid-cycle LH surge.
○Masao Jinno, Aiko Watanabe, Jun Hirohama, Naohisa Hatakeyama, Rie Hiura, Rika Nishiyama
Women's Clinic Jinno, Tokyo, Japan

Objective: In the long protocol of GnRH agonist and hMG regimen, desensitization of pituitary prevents premature
LH surge and thus it has been believed that mid-cycle LH surge cannot be induced. Serendipitously, however, wehave found a novel protocol to induce diminished but significant LH surge, increasing dramatically the rates ofclinical and ongoing pregnancies. Designs: a prospective randomized study. Materials and Methods: Hundred
forty-two initial IVF/ICSI cycles in 140 women with age younger than 41 years and day-3-FSH less than 15 IU/L
were assigned prospectively and at random to receive the conventional long protocol (control group) or a novel
protocol (surge group). In both groups, buserelin acetate was administered nasally at 3 times of 300 μg daily frommid-luteal phase in the preceding cycle until 15:00 pm on the day of hCG administration. In the surge group, 300
μg buserelin acetate was additionally administered twice one hour before and just before administration of 10,000
IU hCG at 21:00 to 24:00 pm, while hCG alone with no additional buserelin acetate was administered in thecontrol group. In 25 patients, concentrations of LH, FSH, PRL, TSH, GH, hCG, E2 and P4 in serum were
measured one hour before, just before and one hour after hCG administration. Results: The rates of clinical andongoing pregnancies per stimulated cycle were significantly (P<0.01 and P<0.05, respectively) higher in surge
group (41% and 34% in 71 cycles) than control (18% and 15% in 71 cycles). In surge group 86% and 14% of 14
hormone-measured women showed increase and plateau, respectively, in serum LH levels after administration of
buserelin and hCG, while all of 11 hormone-measured women in control group showed decrease in LH after hCG(incidence of LH surge: 86% vs. 0%, P<0.0001). The profiles of GH and P4 changes around hCG administration
also significantly differed between surge and control groups. Conclusions: Under pituitary desensitization by nasal
administration of GnRH agonist, diminishied but significant LH surge can be induced by administration of GnRH
agonist at increased dosage within one hour before hCG. This novel protocol significantly improved embryonicdevelopment and ongoing pregnancy rate, suggesting critical role of mid-cycle surge of LH and other hormones inoocyte maturation.

O-028

GnRHa: a promising alternative of HCG to trigger ovum maturation in minimal
stimulation cycle
Ze Wu, Bo Deng, Yonggang Li, Yunxiu Li, ○Yanping li
Department of Reproduction and Genetics; Reproductive Medicine Centre, The First Peoples Hospital of Yunnan Province, China

The objective of this study was to assess the effect of recombinant HCG and GnRH agonist to trigger ovum
maturation in minimal stimulation cycle. This comparative study, at the First People's Hospital of Yunnan
Province, was performed on 189 infertile IVF embryo transfer candidates. Inclusion criteria were normal basal
hormonal profile, <lt38 years old of age, cause of infertility were male factor, tuble abnormalities. A total of 189
infertile patients undergoing IVF/intracytoplasmic sperm injection (ICSI) treatmen with Clomiphene Citrate(CC)/human menopausal gonadotrophin (HMG) were prospectively studied. All the cycles were gouped according
to two different ovum maturation trigger agents: recombinant human chorionic gonadotrophin (rec-HCG) or
GnRHa. Oocyte retrieval was performed 32-34 h after reHCG or GnRHa injection subcutaneously. One to threeembryos were transferred on day 3. The principal outcome wes to evaluste clinical pregnancy and embryo
implantation rates. The two groups were similar in terms of patients' age, day 2 FSH and LH, number of retrived
oocyte, oocyte retrival rate, fertilization rate, cleavage rate and number of transferred embryo. In contrast, thepregnancy rate (13.0 versus 25.6%) was better in the GnRHa group, but there was no signicant difference between
the two groups. The implantation rate was significantly higher in the GnRHa group (25.42 versus 10.45%,
p<lt0.05) compared with rec- HCG group. In conclusion, GnRHa seemed to be a promising and better alternativeof HCG to trigger alternative of HCG to trigger ovum maturation in minimal stimulation IVF cycle.

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●Oral Communications

O-029

The observation of effect of two different COH protocols on the IVF-ET outcome ofaged patientsBo Deng, Ze Wu, Yanping Ma, ○Yonggang Li
Department of Reproduction and Genetics; Reproductive Medicine Centre, The First Peoples Hospital of Yunnan Province, China

To explore the influence of two different COH protocols on IVF-ET outcome, and the better COH protocols foraged infertility. Two hundred infertility patients who were 35 -45 years old received in vitro fertilization andembryo transfer ( IVF-ET ) from May 2010 to May 2011 were randomly divided into two groups, the patients ingroup A accepted the GnRHa+ Gn short protocol while the patients in group B accepted the GnRH ant+ Gn
protocol. Duration of stimulation, amps of gonadotropin, estradiol level on hCG day, number of oocytes retrieved,
fertilization rate, total embryos obtained, high quality embryo obtained, embryos transferred, embryos
frozen,implantation rate per transfer,clinical pregnancy rate per transfe, miscarriage rate between A, B groups werecompared. The level of serum estrogen (E2) on the HCG injecting day and the number of oocytes retrieved ofgroup B were less than that of group A. The duration of stimulation, dosage of Gn of group B were higher than thatof group A, but there was no statistical significance. There were no significant difference in the fertility rate, thehigh quality embryos rate, abortion rates between the two groups.The clinical pregnancy rate in group B was higher
than those in group A (35% vs 21%), there was statistical significance between two groups (P<0.05). Although the
estradiol level on hCG day and the number of oocytes retrieved are lower in the GnRH-ant protocol, which does
not affect the implantation rate and clinical pregnancy for aged infertility, the GnRHant protocol is excelled which
can have better IVF-ET outcome.

O-030

EFFICACY OF SEQUENTIAL TREATMENT PROTOCOL WITH HIGHLY
PURIFIED URINARY FSH AND RECOMBINANT FSH FOR CONTROLLEDOVARIAN STIMULATION○Hong Ye, Guoning Huang, Li Pei, Pinghong Zeng, Xiu Luo
1

Chongqing Genetic and Reproductive Institute, Chongqing Obstetrics and Gynecology Hospital, China

OBJECTIVE: To evaluate impact of sequential stimulation protocol with different FSH isoforms on oocyte quality
and IVF/ICSI outcomes. DESIGN: A prospective randomized, open-label, controlled study. MATERIAL AND
METHODS: A total of 204 IVF/ICSI cycles were included in this study between June 2010 and January 2011. The
inclusion criteria were (1) age between 25-34 years old; (2) BMI between 18-25 kg/m2; (3) First IVF/ICSI; (4)normal ovulatory cycles; (5) both ovaries present and normal uterus; (6) infertility attributable to tubal, male, or
idiopathic factors. The participants underwent a luteal phase gonadotropin releasing hormone agonist
down-regulation protocol. After pituitary down-regulation was confirmed, the patients were randomized into threegroups according to computer-generated random numbers: group A (n=67) received recombinant FSH, (r-FSH
Gonal-F, Serono, contains more less-acidic FSH isoform) only until hCG administration; group B (n=72) received
highly purified urinary FSH (HP-FSH Fostimon IBSA, contains more acidic FSH isoform) only until hCGadministration; group C (n=65) sequentially treated HP-FSH for first 5 days and followed r-FSH until hCGadministration. In three groups, FSH starting doses 150 IU/day for first 5 days and then adjusted according toovarian response. IVF/ICSI outcomes among three groups were compared. RESULTS: There was no significant
difference for duration and cause of infertility, age of patients and type of IVF/ICSI procedure, doses of totalgonandotropin and days of stimulation as well as number of embryo transfer. CONCLUSION: ovarian stimulation
seems to be affected by different FSH isoforms. The sequential protocol improves oocyte maturity and embryocleavage, and increases pregnancy compared with stimulation with acidic HP-FSH alone.

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●Oral Communications

O-031

Application of elective single embryo transfer (e-SET)○Yonggang Li, Yanping Ma, Ze Wu, Lian Deng, Bo Deng, Mengying Gao
Dep of Reproduction and Genetics, Yunnan first people hospital, Kunming, China

Introduction: Elective single embryo transfer (e-SET) is now considered to be one optional way to solve multiple
gestations. The objective of this report is to discuss strategies of SET. Material & Methods: From Jul.2009 to
Dec.2010, We performed 1380 IVF cycles with infertility cause of tubal factor, including 4 groups: 1-SET of 4-8
cells (168 fresh and 46 frozen cycles), 2-single frozen blastocyst transfer (SBT,98 cycles), 3-multi-embryo transferof 4-8 cells(794 fresh and 164 frozen cycles), 4-multi-blastocyst transfer (110 cycles). The relationship between
multiple factors (age of patients, COH protocol, embryo quality, et al.) and clinical pregnancy rates (PRs) as wellas multiple gestation rates were statistically analyzed. Results: 1.PRs achieved 24.3% in group 1, which wassignificantly lower than in group 2 (42.8%), group 3 (42.9%), and group 4(53.6%). PRs were obviously lower ingroup 1 (24.9% of fresh, 23.7% of frozen) compared with group 3 (43.6% of fresh, 39.6% of frozen). 2.Age: PRs
had no notable differences between younger patients(<35) and elder ones(>35) in group 1 (26.4% to 22.1%), group
2 (43.6% to 41.5%). Minimal difference appeared in group 3 (45.2% to 39.1%). But in group 4, PRs of younger
ones were higher than that of older ones (58.2% to 40.7%). 3. COH protocols: In group 2, PRs of low stimulation
reached 56.8%. In group 4, PRs were significantly higher in low stimulation group (58.4%) than in others. 4.
Embryo quality: In group 2, PRs of transferring high-quality blastocysts (52.7%) were significantly higher thantransferring low-quality ones (39.4%). PRs of transferring 2 high-quality ones (59.4%) were slightly higher thantransferring 1 high-quality plus 1 low-quality blastocysts(48.4%), as well as transferring 2 low-quality ones
(47.2%). But in group 1 and 3 the influences were not so obvious. 5. Multiple gestation rates: Multiple gestation
rates were obviously lower in SET group (0%) than in multiple transfer group.Conclusions: Though PRs ofmulti-embryo transfer are higher than that of SET (both fresh and frozen), frozen SBT can obtain equal PRs as well
as frozen multi-blastocyst transfer while patients are over 35 years old. Besides, PRs of high-quality frozen SBT istwice than that of low-quality frozen SBT. Based on the data, frozen SBT, especially high-quality frozen SBT, hasbeen demonstrated to be a valuable choice for elder patients to reduce multiple gestations.

O-032

A Novel function of Human Pinopodes by Expressing L-Selectin Ligand during
Window of Implantation
1

2

3

4

○M Kabir-Salmani , R Nejatbakhsh , H Hosseini , E Dimitriadis , M Iwashita
1

52

National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran., Dept. Anatomical Sciences, Shaheed Beheshti
Medical University, Tehran, Iran, 3Molecular and Cellular Biology Research Center, Shaheed Beheshti Medical University, Tehran,Iran, 4Prince Henry's Institute of Medical Research, Australia., 5Dept. Ob/Gyn, Kyorin Medical University, Tokyo, Japan

Background: Apical surfaces of human endometrial epithelium and endothelium are key elements for the initiationof molecular interactions to capture the blastocyst or leukocyte, respectively. The L-selectin adhesion system has
been strongly proposed to play an important role in the initial steps of trophoblast adhesion and promotion of
integrin-dependent processes, ultimately culminating in the establishment of the embryo-maternal interface. On thebasis of these facts, we hypothesized a novel role for uterodomes as the first embryo-fetal contact sites to containthe highest subcellular expression of L-selectin ligand suggesting its role in early adhesion as predicted. Thus, theobjective of this study was therefore to determine the subcelluar pattern of distribution of the L-selectin ligand(MECA-79) in human endometrial apical membrane region during the window of implantation.Materials andMethods: Endometrial biopsies of 5 fertile females ranging in age between 21 and 36 years in the midluteal phase,that is, days 20-24 of a regular menstrual cycle, were studied using several approaches, including scanning electron
microscopy (SEM), immunostaining for light microscopy and transmission electron microscopy (TEM), andimmunoblotting as well as statistical analysis of the area-related numerical densities of immunoreactive
MECA-79-bound nanogolds to detect the expression pattern and the subcellular distribution pattern of L-selectin
ligand (MECA-79) in human endometrium during the window of implantation.Results: The endometrial biopsies
were scored according the dating criteria of Noyes et al. by an experienced histologist. The SEM images of the
midluteal phase specimens revealed that fully developed uterodomes were abundant in our samples. DAB
immunostaining and immunoflouresent staining as well as immunoblotting revealed that MECA-79 was expressedin the midluteal phase specimens. The results of immunogold TEM illustrated the expression of MECA-79 inhuman uterodomes in the midluteal phase and a higher area-relate numerical density in uterodomes compared tothat of the uterodome-free areas. Conclusions: This is the first demonstration of the molecular localization of
MECA-79 in the human uterodomes which may indicate a novel role for uterodomes to be capable ofshear-stress-dependent tethering-type adhesion in the initial phases of human embryo implantation.

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O-033

Obstetric outcomes of ART pregnancies in women over 35 years old○Xiaohui Tang, Tomoko Adachi, Setsuko Nakayama, Yoshiharu Takeda, Hideki Sakamoto,
Masao Nakabayashi
Department of Obstetrics and Gynecology, Aiiku Maternal and Child Health Center, Aiiku Hospital, Tokyo Japan[Objective] In late years, assisted reproductive technology (ART) has progressed remarkably, and pregnancies by
ART are increasing. On the other hand, detailed analysis of the obstetric outcomes of ART pregnancies is stillinsufficient. For appropriate perinatal management, we studied obstetric complications in women over 35 years old
who conceived by ART. [Methods] The study enrolled 418 women over 35 years old conceived by ART. As acontrol group, 3,517 women conceived spontaneously and delivered at our hospital from January 2005 to
December 2010 were used. The obstetrical complications were compared with this control. [Results] The ratio of
ART pregnancies in women over 35 years old showed increasing trend of approximately 12%. The twin-birth rate
decreased sharply, but it was still approximately 5 times higher than in spontaneous pregnancies. The meanmaternal age and the number of primiparous pregnancy were higher in ART pregnancy (39.1±2.8 years old,
82.6%) than in spontaneous pregnancy (37.4±2.2 years old, 37.7%). The incidence of premature birth (before 34
weeks of gestation), PIH, and placenta accreta were also significantly higher (8.6% vs. 2.6%, 8.9% vs. 4.2%, 2.6%
vs. 0.9%, respectively). If limited for the singleton pregnancies, incidence of placenta previa was also significantly
higher (2.5% vs. 1.2%) in addition to premature birth, PIH and placenta accreta (6.2% vs. 2.4%, 6.7% vs. 4.2%,
3.1% vs. 0.9%, respectively). The incidence of the placenta accrete was significantly higher in women over 40
years old than that in women 35-39 years old among ART pregnancies regardless of singleton or multiple gestation(5% versus 0.8%). Uterine disease was a risk factor of the placenta accreta (OR 2.7, IC1.2-6.05) other than theART (OR 2.44, IC 1.15-5.14). [Conclusion] Incidence of premature birth before 34 weeks, PIH and the placental
disorder are significantly higher in the ART pregnancies than that in the spontaneous pregnancies over 35 years
old. Furthermore, incidence of the placenta accreta was particularly high in the ART pregnancies over 40 years old
thus careful management is required.

O-034

The age of 35 years is the critical age for successful TESE-ICSI in nonobstructive
azoospermic patients with normal karyotype and nonmosaic Klinefelter syndrome.
1

1

2,3

2,3

3,4

○Hiroshi Okada , Yoshitomo Kobori , Mitsunobu Koshida , Ken-Ichi Tatsumi , Kazutaka Terai ,5
Osamu Maruyama
1

Department of Urology, Dokkyo Medical University Koshigaya Hospital, Saitama, Japan, 2Koshida Clinic, 3Umegaoka Women'sClinic, 4Department of Urology, Shakaihoken Kamata General Hospital, 5Department of Urology, Juntendo University Faculty of
Medicine

Introduction: It was reported that the sperm retrieval rates of MD-TESE (microdissection testicular sperm
extraction) declined with age in nonmosaic Klinefelter syndrome (KFS) patients. However, effect of aging of KFS
on fertility rate and pregnancy outcomes is unknown. We retrospectively reviewed our experience of men withnonobstructive azoospermia (NOA) treated with TESE-ICSI over 5-year period. We analyzed fertilization andpregnancy rates in patients with nonmosaic KFS comparing those in NOA patients with normal karyotype treatedin the same period of time. Material & methods: Between 2004 and 2009 a total of 164 patients with NOA (118with normal karyotype; 46 with KFS) underwent successful MD-TESE by a single surgeon. All sperm retrieved
from testes were cryopreserved until ICSI. All patients were followed at least one year after the last embryotransfer. Patients backgrounds, fertilization rate, and pregnancy outcome were compared. The Mann-Whitney U
test was used to compare variables between two groups. Comparison of fertilization rate, pregnancy rate andabortion rate between two groups (NOA patients with normal karyotype and nonmosaic KFS; 35 years old andmore than 35 years old) were done by χ2 test. Results: Serum concentrations of FSH were significantly higher innonmosaic KFS than in NOA patients with normal karyotype. However, serum concentrations of FSH weresignificantly lower in those younger than 35 years of age than in those older than 35 years of age in both groups.
Serum concentrations of testosterone were significantly lower in nonmosaic KFS than in NOA patients withnormal karyotype. However, serum concentrations of testosterone were higher in those younger than 35 years ofage than in those older than 35 years of age in both groups. Fertilization rates and clinical pregnancy rates weresignificantly higher in nonmosaic KFS than in NOA patients with normal karyotype. However, fertilization rates
and clinical pregnancy rates decreased significantly over 35 years of age in both groups. Conclusions: Sperm fromtestes of nonmosaic KFS had better fertilizing ability than those from NOA patients with normal karyotype.
However, sperm fertilizing ability decreased after the age of 35 years of old in both groups.

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O-035

Comparing study of effectiveness of nifedipine and magnesium sulfate for acutetocolysis of preterm labor and threatened preterm labor
1

2

3

4

○Fariba Nanbakhsh , Farzaneh Broomand , Zahra Yekta , Rita Doosti , Pooya Mazloomi

51Urmia University of Medical Sciences,Urmia,IRAN, 2Urmia University of Medical Sciences,Urmia,IRAN, 3Urmia University of
Medical Sciences,Urmia,IRAN, 4Urmia University of Medical Sciences,Urmia,IRAN, 5Urmia University of Medical
Sciences,Urmia,IRAN

Introduction: Preterm labor with 10% prevalence is one of the most important complications in newborns withprematurity and makes problems emotionally and economically especially in families with long history ofinfertility. Preterm labor is described with dilatation more than 1cm, effacement more than 85% and threatened
preterm labor is defined as occurrence of contractions without dilatation fewer than 37 th weeks of pregnancy.
Therefore all gynecologists try to find the best method to control preterm labor. All medications used in preterm
labor have some complications. Because of that, we aimed to use nifedipine with most beneficence and least
complications and cost. Materials and Methods: This clinical trial study was performed on 220 patients withpreterm- labor and threatened preterm labor during 2009-2010 years in kosar Hospital of Urmia University of
Medical Sciences. Patients divided in to two groups after description treatment to them and getting informed
consent. Within each group 55 patients were treated with Nifedipine orally and 55 patients were treated withmagnesium sulfate intravenously. Data were collected based on patients response to medications and analyzed with
Spss software. Results: In this study the effectiveness of nifedipine in preterm labor group was more thanMg-sulfate but not statistically significant and in threatened preterm labor group the effectiveness was higher
significantly (p=0.001). Also the mean time from treatment to delivery was significantly more in nifedipine group.
(PV=0.000) Nifedipine group had less maternal complications and better fetal outcomes significantly. (PV=0.000)
Conclusion: These findings suggest using nifedipine would be considerable for prevention of preterm labor andthreatened preterm labor.

O-036

The Change of Serum Anti-Mullerian Hormone (AMH) Level by Chemotherapy andOperation in Premenopausal Women with Gynecological Cancer○Masaru Hayashi, Akiko Shoda, Nobuaki Kosaka, Yoshiko Moshiduki, Ichio Fuykasawa
Department of Obstetrics and Gynecology, Dokkyo Medical University, Tochigi, Japan

Objectives: Women who underwent chemotherapy (chemo) sometimes resulted in premature ovarian failure (POF).
Recently, the serum AMH level has been shown to be a marker of ovarian reserve. The objectives of this study are(1) to observe the change of serum AMH level as a marker of ovarian reserve by chemo and operation (ope) for
patients with gynecological cancer, (2) to evaluate the pathological finding of the ovary after chemo in the AMH
measured cases. Subjects: 5 premenopausal women, who underwent chemo with or without ope against
gynecological cancer in our department, were studied. Mean age of them was 28.4 (range 13-45) years, and they
were two cases of ovarian germ cell tumor (GT), one case of epithelial ovarian cancer (OC), one case of corpus
uteri cancer (CUC) and one origin-unknown case of cancer pleuritis and peritonitis (CPP). The performed chemo
regimens were BLM+VP-16+CDDP for GT, DTX+CDDP for CUC, TC (PTX+CBDCA) for OC, three regimens;
TC, CPA+ADM+CDDP and PLD for CPP. Design and Methods: Informed consents were obtained from patientsin this study. Serum AMH, LH, FSH, and E2 level were measured before, during, and after chemo and ope. AMH
level was measured ourselves using AMH immunoassay kit (Immunotech).Results: (1) Serum AMH level of allcases were gradually decreasing according as chemo and it could not be detected after the second or third course.
(2) In two GT cases, who were in her teens and twenties, serum AMH level re-increased in a few months after the
final course of chemo. Afterward, their AMH value reached to the low plateau level as compared with the same
aged women. They have regular menstrual cycles, and normal serum gonadtropin-E2 levels. However, theirovarian reserves are presumed to be impaired and the almost equivalent level with thirties' and forties' level interms of AMH level. (3) The pathological study revealed that (a) ovarian follicles were degenerated, decreased
their number, and impaired their growth, (b) the small blood vessels and fibrosis in the cortex were proliferated.
Conclusion: (1) Even if they have normal menstrual cycle, and normal gonadtropin-E2 levels, the ovarian reserves
are impaired by chemo and ope. It is seemed to be able to predict it by measuring the serum AMH. (2) The patient
who shows hypergonadtropic-hypogonadism is not always POF during and right after chemo. We should evaluate
their ovarian reserve after some periods in terms of ovarian follicle development.

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O-037

Alternative treatment options for patients with ovarian insufficiency basedon
Natural Cycle IVF○Markus Nitzschke
Milagro Kinderwunschzentrum Bodensee AG, Kreuzlingen, Switzerland

Introduction: In general, patients with low ovarian reserve are difficult to manage and have a relatively poor
prognosis. Using traditional stimulation protocols like the long GnRH agonist protocol or the GnRH antagonist
protocol in these patients often results in low ovarian response and poor pregnancy rates. In natural menstrual
cycles without ovarian stimulation, every month the female body naturally selects the best possible oocyte forovulation. Nevertheless, hormonal changes due to ovarian insufficiency can influence and change the individual
menstrual cycle pattern of each patient over the time, which may result in difficulties to conceive naturally. We
observed the menstrual cycle pattern of patients with low ovarian reserve in order to distinguish different stages ofovarian insufficiency. Once we were able to describe the different stages, we developed new treatment approaches
for each patient group based on Natural Cycle IVF. Material & Methods: In 2010, the menstrual cycle pattern of 10
patients with AMH <1.0 nmol/L were observed. Patients were 22 to 42 years old. Blood samples were drawn todetermine FSH, LH, E2 and transvaginal ultrasound scans were performed on different days of the cycle.
Depending on the cycle pattern of each patient, we offered individualized treatment approaches based on Natural
Cycle IVF using either Ibuprofen or Clomifen to control ovulation, GnRH agonists to induce ovulation and either
Ethinyl-Estradiol or combined oral contraceptive pills to regulate the cycle. Fresh embryo transfer was performedon day 2 or embryos were frozen and transferred later in artificial cycles. Patients were informed about off label
use of the medication and informed consents were signed. Results: Based on our observation, we could describe
four different stages of ovarian insufficiency. We were able to perform oocyte retrievals and embryo transfers in all
10 patients. A total of 33 natural cycles were initiated. Premature ovulation occurred in 3 cycles (9.0%) and no
retrieval was attempted. Among the attempted 30 oocyte retrievals, 21 (70.0%) were successful. Out of those 21
oocytes 11 (36.6% per retrieval) were mature and 10 (33.3% per retrieval) were immature. ICSI resulted in 8
fertilizations (72.7% per mature oocyte). Out of 8 transfers, 3 (37.5%) resulted in biochemical pregnancy. Twopatients delivered (25.0%), one patient had a miscarriage at 8 weeks of pregnancy. Conclusion: Our experience
shows that ovulation can successfully be controlled by the use of NAIDs or clomiphene citrate and does notnecessarily require GnRH analogues for pituitary suppression. This knowledge opens new space for developmentof alternative protocols respecting the patients‘ own physiology with no need for heavy stimulation. Patients withovarian insufficiency may benefit from this approach, which can be offered before referring them to egg donation.

O-038

CORRELATION OF BODY MASS INDEX WITH OUTCOME OF IN VITROFERTILIZATION IN A DEVELOPING COUNTRY○Neeta Singh, Prerna Gupta, Suneeta Mittal, Neena Malhotra, Anupama Bahadur
All India Institute of Medical Sciences, New Delhi, India

Introduction: To correlate ovarian response to stimulation and IVF outcome according to the body mass index(BMI). Materials and Methods: We retrospectively reviewed all patients who underwent IVF cycle in our
institution from January 2008 to October 2010. Three hundred and twenty eight patients underwent 342 in vitro
fertilization cycles or ICSI and were divided into four subgroups according to BMI; underweight BMI; 18.5kg/m2;
normal weight BMI-18.5-24.9; overweight BMI-25.0-29.9 and obese more than 30.0 kg/m2. Results: Three
hundred and forty two in vitro fertilization cycles or ICSI from our IVF database were studied. According toclassification by BMI, 8.8% of women were underweight (BMI; 18.5 kg/m2), 43.3% were normal weight (BMI
18.5-24.9) 41.2% were overweight (BMI 25.0-29.9) and 6.1% were obese (BMI more than 30.0 kg/m2). The totaldays of stimulation was almost similar, approximately 10-11 days but total dose of gonadotropin required,increased as BMI increased (p value 0.04). We found that with increasing BMI negative co-relation was seen withclinical pregnancy rate (p value=0.40) with pregnancy rate decreasing from 31.2% in normal weight woman to
21.9% in obese women. In the present study no difference was seen in the number of oocyte retrieved but adecreased fertilization (75.8% in normal weight to 66.1% in obese) and cleavage rate (72% to 61%) was seen withdecreased number of cryo-preserved embryos (4.32 to 2.4) with increasing BMI. This study shows that poorer
oocyte quality is seen with increasing BMI which results in reduced clinical pregnancy rate. In our study nodeleterious effect of low BMI was seen on IVF outcome and clinical pregnancy rate. Conclusion Female obesity
impairs IVF outcome potentially by impairing oocyte quality but does not affect ovarian response to stimulation.

257

●Oral Communications

O-039

Bilateral ovarian endometriomas removal does not affect the IVF outcome○Andrej Vogler, Martina Ribic Pucelj, Irma Virant Klun
Department of Obstetrics and Gynaecology, University Medical Centre, Ljubljana, Slovenia

Introduction: Endometriosis represents nowadays probably the most frequent cause of female infertility or
subfertility. At our institution endometriosis is diagnosed in more than 40 % of infertile patients. Treatment ofchoice is by all means laparoscopic surgery which leads to a more than 60 % pregnancy rate, regardless the stage ofthe disease. For the rest of the patients in vitro fertilization (IVF) is the most appropriate treatment of choice. The
impact of ovarian endometriomas on IVF outcome remains still controversial and is a matter of debate. Whereassome studies have documented that ovarian endometriosis is associated with a reduced ovarian reserve andconsequently lower pregnancy rates, others failed to prove this association. The present study was designed toelucidate if bilateral ovarian endometriomas removal whether or not affects IVF outcome. Materials and methods:
In 34 patients (group A) underwent IVF program the only cause of infertility was bilateral ovarian endometriomas
which had been previously surgically removed. The control group (group B) represents 168 patients who
underwent IVF procedure due to tubal factor. The inclusion criteria for the study were age less than 37 years,
regular ovulatory cycles, normal basal gonadotrophin level and normal spermiogram. Among others, following
parameters were analyzed and compared between the both groups: fertilization rate, pregnancy rate per cycle andper embryo transfer (ET), take home baby rate (THBR) per cycle and per ET. Results: In none of the comparedparameters statistically significance was reached. In the group "A" pregnancy rate per cycle and per ET was 44.1%
and 55.6 % compared to the group "B" where was 38.7 % and 42.5 % respectively. THBR per cycle and per ET inthe group "A" was 38.2 % and 48.1 %, whereas in the control group was 33.3 % and 36.6 % respectively.
Conclusions: Severe ovarian damage may occur in ovaries operated for ovarian endometriomas but the present
study showed that bilateral endometriomas removal does not affect IVF outcome in our patients. Pregnancy rateand THBR were even higher, though not statistically significant, than in patients with tubal factor of infertility. We
believe that appropriate surgical technique is of paramount importance to preserve functional ovarian tissue inpatients with endometriomas leading to favourable IVF outcome if needed.

O-040

The new stripping technique for low protrusion rate uterine myoma in TCR○Toshimichi Oki, Chie Oki, Toshihiko Kawamura, Akiko Gibo, Hideki Yamasaki, Tsutomu Douchi
Wemen's Medical Center, Kagoshima Univesity Hospital, Kagoshima, Japan[Objective] In present study, we studied the effectiveness of the new stripping technique in TCR (transcervicalresection) for low protrusion rate uterine myoma. [Design] Four cases had low protrusion rate (less than 40%) ofuterine submucosal myoma. Before TCR, cervical dilation was done with more than 5 pieces of laminalia. Our new
stripping technique of myoma is shown as follows. At first, transverse incision was made on the top of myoma
nodule. The tip of a loop electrode was inserted to the space between myoma nodule and the muscle layer of uterus
without electrocagulation, and myoma nodule was detached from normal layer of uterine muscle. Until the
protrusion rate reach more than 70% and diameter of myoma reach to less than 2cm, resection and detachment arecontinued. [Results] In four case, TCR was successful without complication. [Conclusion]The new stripping
technique in TCR has contributed to success TCR for low protrusion rate uterine myoma.

258

●Oral Communications

O-041

Evaluation of short-term and long-term outcome of tubal conservation in thetreatment of tubal ectopic pregnancy○Toshimichi Oki, Yukiko Nakajou, Chie Oki, Akiko Gibo, Toshihiko Kawamura, Tsutomu Douchi
Wemen's Medical Center, Kagoshima Univesity Hospital, Kagoshima, JapanOBJECTIVES: To evaluate short-term and long-term outcomes of tubal conservation in the treatment of tubal
ectopic pregnancy such as laparoscopic salpingostomy (LS) and methotrexate (MTX) treatment. We assessed interms of consequent tubal preservation and tubal patency for the short-term outcome, and pregnancy rate andectopic pregnancy rate for the long-term outcome. METHODS: This study include 131 cases who underwent LS(LS group) and 21 who were treated with systemic administration of MTX (MTX group) among 216 ectopic
pregnancy cases treated in the past 8 years. Serum human chorionic gonadotrophin (hCG) concentration andChlamydia trachomatis IgG and IgA antibodies were measured before the treatment, and serum hCG concentration
was measured until it turned to negative after the treatment. Four months after the treatment tubal patency wasassessed by using hysterosalpingography or hydrotubation (113 of LS group and 13 of MTX group). We evaluatedthe long-term outcome of the cases (57 of LS group and 9 of MTX group) desiring future pregnancy and were able
to follow over a year. RESULTS: The short-term outcome of LS group / MTX group was as follows; tubalpreservation rate: 93.1% (122/131) / 95.2% (20/21), tubal patency rate: 96.5% (109/113) / 84.6% (11/13). There
was no significant difference between the groups, and these rates were not affected by serum hCG concentration
before the treatment, C. trachomatis antibodies or fetal cardiac activity. Also, no difference was detected in thelong-term outcome of LS group /MTX group; intrauterine pregnancy rate: 26.3% (15/57) / 33.3% (3/9), ectopic
pregnancy rate: 8.8% (5/57) / 0% (0/9). However, it was distinctive of five recurrent ectopic pregnancy cases that
all of them were positive for C. trachomatis antibodies and were detected peritubal adhesion at the first surgery andthat four of them were ipsilateral tubal ectopic pregnancy.

O-042

2000IU hCG in high responders does not affect the outcomes of IVF
1

1

1○YanPing Kuang , Yun Wang , QiFeng Lyv , John Zhang

2

Department of Assisted Reproductive, Shanghai Ninth People‘s Hospital Affiliated Shanghai JiaoTong University School of Medicine,
Shanghai, China, 2 New Hope Fertility Center, New York, U.S.
1

To evaluate the effect of 2000IU and 5000IU hCG in high responders during IVF-ET. One hundred and eighty-two
IVF cycles were analyzed from high responders based on oocyte quantities, embryo transferred times and peak E2
levels. On the day of hCG administration, if E2 levels were ≥3000 but <5000pg/ml, patients received 5000IU(group B). For E2 levels≥ 5000 pg/ml, patients received 2000IU (group A and C). If patients failed to get pregnant
after they transferred embryos≥3 times, we cryopreservated all their embryos after oocyte retrieval (group C).
Mean ages were 30.18 ±3.19, 30.33±3.29 and 29±2.63 for groups A, B and C. Peak E2 levels in group A or C washigher than group B (6788±2975, 5597±2240vs. 4630±815 pg/ml), as well as the mean number of oocytes
retrieved (21.53±7.4, 22.89±8.71vs. 18.34±4.84). Proportion of mature oocytes (90.56%, 89.45%vs. 87.65%),
fertilization rates (71.97%, 74.4%vs. 74.07%), high qualityrates (66.16%, 61.87%vs. 58.23%), chemical PR(44.83% vs. 44.83%) and clinical PR were similar. No incidence of moderate or severe OHSS was found in 2000
IU hCG, but two moderate OHSS patients were found in 5000IU hCG group. Taken together, 2000IU hCG is safeand efficient to induce adequate oocyte maturation, fertilization while eliminating the risk of OHSS in highresponder patients.

259

POSTER PRESENTATIONS

260

●Poster Presentations

P-001

The influence of crowding on Leydig cell, weight gain, testosterone and cortisolelevels in mice
○Maryam Ghasemi, Farzad Rajaei
Department of Anatomy, Qazvin University of Medical Sciences, Qazvin, Iran

Aims: Human typically encounter in the modern, developed world with stressors that originates largely from social
and interpersonal interactions. Epidemiological data suggest that stress is related to the development of metabolic
disease and infertility. In the present study the effects of crowding on Leydig cell, cortisole and androgen levels
were investigated. Methods: 60 male mice were divided into 6 groups. Control groups (2 cages, 5 mice/cage),groups 1 as mild stress groups (2 cages, 10 mice/cage) and groups 2 as severe stress (2 cages 15 mice/cage). Out ofsix groups, three groups kept for one month and three next groups kept for two months. The mice wereanaesthetized with an intra peritoneal injection of ketamine and xylazine after one and two months. The
testosterone and cortisol levels in plasma were assessed by radioimmunoassay. Then, the samples of testes were
removed and processed for light and electron microscopy. The data has been compared using statistical methods(SPSS, ANOVA, P<0.05). Results: The results showed that the differences in weight gain and plasma testosterone
levels in animals exposed to stress for one month were not significant as compared with control group (P<0.01);
Whereas the differences in the number of leydig cells in groups 3 were significantly increased as compared with
control group (P<0.01). The differences in other study groups weren't significant. The plasma cortisol levels ingroup 2 were significantly increased as compared with control group (P<0.02) and the plasma cortisol levels ingroup 3 were also significantly decreased as compared with control group (P<0.006); whereas the differences inother groups weren't significant. The results showed that the differences in the numbers of leydig cells, weight
gain, plasma testosterone and cortisol levels, in animals exposed to stress for two months weren't significant.
Transmission electron microscopy revealed decreasing the numbers of secretory granules in the Leydig cells inmice of group 2 and 3 as compared with control group. Conclusion: These results showed that crowding stress canaffect on number of leydig cells and plasma cortisol levels on male mice.

P-002

Identification of basic reprogramming factors associated with a pluripotent potential
of in vitro cultured spermatic stem cells in domestic animals.
1

1,2

3

1

1

Sung-Min Kim , Mayako Fujihara , Sadeep Goel , Mahesh Sahare , Naojiro Minami ,1
1
Masayasu Yamada , ○Hiroshi Imai
1

Kyoto University, Kyoto, Japan, 2Smithsonian Coservation Biology Institute, Front Royal, Virginia, USA,
Center for Cellular and Molecular Biology, Hyderabad, India

3

Spermatogonial stem cells (SSCs) that are present in the mouse testis have the ability to give rise to pluripotent
ES-like cells under in vitro conditions. However, properties of the potential have poorly understood in domestic
animal species. Transcription factors POU5F1, SOX2, c-MYC, KLF4 and NANOG have been known to be essential
for maintaining pluripotency of stem cells and generation of iPS cells. In the present study, we have identified
these reprogramming factors in the testis of domestic animals. Spermatogonial stem cells/gonocytes in both pig andbull testis expressed UCHL-1 and VASA, which are known as germ cell specific markers in both species.
Interestingly, germ cells that are express UCHL-1 and VASA simultaneously expressed POU5F1 and NANOG
protein, and the expressions of SOX2, c-MYC and KLF4 genes were also observed. Under in vitro cultivation ofstem/progenitor spermatogonia, NANOG and POU5F1 protein were specifically expressed and ES cells-likecolonies were formed together with the expressions of germ-cell specific markers. In addition, spermatic stem cells
differentiated into various cell lineages including three germ layers. These results suggest that unipotent germ stem
cells in domestic animals have a reprogramming potential into pluripotent stem cells in in vitro culture condition.

261

●Poster Presentations

P-003

Effect of diet contain sesame seed on the rat testis
121Javad Amini mohabadi , ○Hassan Hassani Bafrani , Morad Pasha Eskandari Nasab ,1
3
4
Mohammad Hossein Shahir , Hossein Nikzad , Aliakbar Taherian
1

Zanjan University, 2Anatomical Research Center, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran,
Department of Anatomy & Embryology, Scientific Director of the IVF Lab, School of Medicine, Kashan University of Medical
Sciences, Kashan, Iran, 4Department of Anatomy & Embryology, Scientific Director of the IVF Lab, School of Medicine, Kashan
University of Medical Sciences, Kashan, Iran
3

Introduction: The development of a new dietary adjunct with a novel natural antioxidant impact on the malefertility in general has been increasingly expressed in recent time. However, the purpose was to evaluate the effects
of sesame seed on adult male rats testis using unbiased. Methods and Materials: Histopathology, biochemical andhormonal studies. Thirty adult male rats were divided into two groups of 15 rats each. The regimen group received
diet containing 30% sesame seed, while the control group received diet along eleven weeks. Five microns of
uniformly random transverse sections of processed testicular tissues were equally analyzed. Testis was analyzed
using Graph pad t test software and P<0.0001 was considered extremely statistically significant.Results: The
results showed that no significant body weight rats, weight and volume testis and percentage volume seminiferous
tubules vessels. However, the mean cells number of epithelial and percentage volume of epithelial, lumen andinterstitial of this tubules were extremely significant (P<0.0001) respectively, in the experimental group compared
to control. Conclusion: Sesame seed intake may improved testicular parameters, fertility and sperm production in
the males.

P-004

Analysis of Y chromosome microdeletions in Kashanian infertile males
1

2

1

3

○Mahnaz Torfeh , Hassan Hassani Bafrani , Ebrahim Sakhinia , Mahdi Rohani
12Department of Biochemistry and genetics, Faculty of Medicine, Tabriz University of Medical Sciences, Anatomical Research Center,
School of Medicine, Kashan University of Medical Sciences, Kashan, Iran. , 3Department of Bacteriology, Pasture Institute of IranIntroduction: Studies have shown that approximately 10-15% of couples have infertility problem and a male factor
can be accounted for half of these cases. Deletion in three non-overlapping regions on long arm of Y chromosome(AZFa, AZFb and AZFc) is caused has defective spermatogenesis and ultimately infertility in men. According torecent studies males who have Y chromosome deletions and followed by ICSI may transmit their deletions to a sonoffspring. So before ICSI all such men should undergo screening of these deletions for preventing transmitting totheir son. Methods & Material: Screening of Y chromosome micro deletions was done in 50 infertile men referring
to kashan infertility center for ART. Genomic DNA was extracted from peripheral blood lymphocytes by standard
ethanol chloroform techniques. PCR performed on each samples for detection of micro deletions, using 8 STS
markers based on EAA/EMQN guideline and 12 STS markers which used in Iran and neighbor countries. Results:
In this study, none of patient has these deletions. Conclusions: This study failed to find any deletion in the patients
who came for ICSI to this centre, so it appears that other factors are important in the infertility of this population.

262

●Poster Presentations

P-005

The usefulness of testicular sperm extraction with the Trucut biopsy needle○Syuichi Iida, Masakuni Suzuki, Ikuo Tachibana, Sigetomo Takahashi, Takahiro Noda, Osamu
Fuzii
M, Suzuki's Memorial Hospital, Miyagi, Japan

Introduction: Currently, testicular sperm extraction (TESE) can be performed in many ways to obtain testicularspermatozoa from azoospermic patients. TESE is mostly performed by invasive surgical procedures under total or
lumbar anesthesia, which are traumatic for patients and also very technically demanding. We performed TESE
under intravenous anesthesia by using the Trucut biopsy needle. This technique is less technically demanding andless invasive than open surgical procedures. Further, this method is advantageous because TESE can be repeated,unlike open surgical procedures. The purpose of this study was to assess the usefulness of TESE performed using
the Trucut biopsy needle. Methods: In this study, the subjects were 105 infertile men who underwent TESE thatwas performed using the Trucut biopsy needle. The relationship of serum follicle stimulating hormone (FSH)levels with the sucess rate of TESE was analyzed. In TESE, the presence or absence of testicular spermatozoa was
examined at first. Subsequently, the motility of the testicular spermatozoa was investigated. Small amount of
testicular tissue was collected for histopathological examination. Results: Testicular spermatozoa were retrieved
from a total of 64 (61%) patients of which 57 (88%) had an FSH level <16 mIU/ml. Among patients in whom
TESE was successful, motile sperms were identified in 44 (69%) of which 43 (98%) had FSH levels <16 mIU/ml.
In 11 (23%) patients with a histopathological diagnosis of germ cell aplasia, testicular spermatozoa were retrieved.
Conclusion: TESE performed using the Trucut biopsy needle is as effective as open surgical procedures in patientswith FSH levels <16 mIU/ml. However, in patients with FSH level >16 mIU/ml, the proportion of patients who
had immotile spermatozoa increased, even if testicular spermatozoa were able to retrieved. Therefore, for patientswith FSH levels >16 mIU/ml, other procedures of TESE are preferred because the possibility of obtaining motile
testicular spermatozoa is higher.

P-006

An evaluation of the confounding effect of sperm abnormalities on pregnancy andimplantation rates and miscarriage rates following in vitro fertilization-embryotransfer using sibling oocytes○Jung K Choe, Jerome H Check, Theresa Jamison
UMDNJ, Robert Wood Johnson Med. School at Camden, Cooper Hosp./Univ. Med. Cntr., Dept. OB/GYN, Div. Repro. Endo. &Infertility, Camden, NJ, U.S.

Introduction: The possibility exists that sperm with certain abnormal parameters may be able to fertilize oocyteswith in vitro fertilization embryo transfer (IVF-ET) and produce normal looking embryos that are inferior and lesslikely to result in a normal pregnancy. The best way to test this hypothesis would be to compare the pregnancy
outcome following the fertilization of oocytes from the same pool of oocytes shared by two males, one with and
one without normal semen parameters. Materials and Methods: Donor oocyte recipient pairs sharing the same poolof donated oocytes were identified where one male partner had a subnormal semen specimen whereas the othermale partners semen specimen was considered normal. To be considered an abnormal specimen the motile density
had to be <10x106/mL, or motility <30% irrespective of sperm concentration or normal morphology using strict
criteria <4%, or the presence of >80% antisperm antibodies or a hypo-osmotic swelling test of <50%. Clinical(gestational sac at 8 weeks) and live delivered pregnancy rates and implantation and spontaneous abortion rates
were compared with these two groups following embryo transfer. Only 1 cycle per patient was used. Many but not
all recipients with husbands with subnormal semen specimens had the oocytes fertilized by intracytoplasmic sperm
injection (ICSI) whereas the majority of the recipients without male factor had conventional oocyte insemination.
Results: There were 138 paired cycles compared. There was no significant difference in fertilization rates for thosewith male factor (785/1088, 72.2%) vs. no male factor (817/1184, 69.0%). There was no difference in clinicalpregnancy rate/transfer 52.2% (72/138) vs. 48.6% (67/138) or live delivered pregnancy rate/transfer 42.0%(58/138) vs. 43.5% (60/138) or implantation rate 29.6% (114/385) vs. 27.8% (109/392) (chi-square analysis). The
miscarriage rate was twice as high with male factor (19.4%, 14/72) vs. 10.4% (7/67) (p=0.16, Fishers exact test).
Conclusions: The presence of male factor in the era of ICSI does not seem to have a negative impact on IVF
outcome as evidenced by comparing outcome of sibling oocytes of which half were fertilized by males with andhalf without subnormal semen parameters. Though the miscarriage rate was twice as high with male factor theslightly higher clinical pregnancy rate per transfer resulted in no difference in the live delivered pregnancy rate per
cycle.

263

●Poster Presentations

P-007

Testicular sperm retrieval using a new multiple-holes puncture needle○Hong-Hua Wang, Li-Yi Cai, Hong-Ying Yu, Jing-Ying Xiang, Lin-Qing Hu, Xiao-Jin Zhou
Department of Reproductive Medicine, the Affiliated Wuxi Hospital for Maternal & Childers Health Care of Nanjing Medical
University, China

Testicular fine needle aspiration (TFNA) was initially used for diagnosis purposes and it is now sometimes used torecover sperm from the testicles for ICSI. This study aimed to compare the reliability of testicular sperm extraction
by a new multiple-hole needle aspiration (MHNA) with TFNA in infertile males with azoospermia. This
retrospective cohort study involved 68 patients with azoospermia. Detailed clinical and laboratory examinations
were performed and two semen analyses were obtained from each patient. A 20-ml 14 gauge 13-mm needle withmultiple-hole was used for MHNA, and a 20-ml 21 gauge 13-mm needle was used for TFNA. The 68 patients
underwent testicular sperm retrieval were divided into two groups according to their sperm retrieval methods(MHNA group n=30; TFNA group n=38). The motile sperm retrieval rate in MHNA group was 70%, significantly
higher than that in TFNA group of 39.5%. Five ICSI cycles were performed in MHNA group, and four cases get
pregnant. Testicular MHNA in infertile males is a simple, reliable and minimally invasive technology for testicularsperm retrieval. It is more effective than TFNA in testicular sperm extraction.

P-008

The impact of body mass index on sperm recovery and serum reproductive
hormone levels in an infertility setting-An analysis of 445 azoospermic cases 1

1

2○Hatsuki Hibi , Tadashi Ohori , Yoshiaki Yamada , Yoshimasa Asada
1

3

2

Kyoritsu General Hospital, Nagoya, Japan, Department of Urology, Aichi Medical University School of Medicine, 3Asada Lady's
Clinic, Aichi, Japan

Introduction: The relationship between metabolic syndrome and health disorders or the decline in male sexualfunctions is a major topic of discussion; moreover, reports describing a correlation between obesity and sperm
count have appeared in the literature. A retrospective study regarding the impact of body mass index on
reproductive hormones and potential sperm recovery was conducted involving infertility cases treated at this clinic
on an outpatient basis over the past decade. Materials and Methods: The subject population consisted of 795
infertility outpatients treated in this clinic between April 2000 and March 2010; 350 and 445 cases of subfertility,e.g., varicocele, and azoospermia (obstructive azoospermia= OA, 187 cases; non- obstructive azoospermia= NOA,258), respectively, were diagnosed. Additionally, 30 fertile controls were included. Levels of reproductive
hormones, triglyceride, HDL-cholesterol and uric acid and BMI were examined. Results: Total and freetestosterone levels displayed significant elevation in NOA sperm recoverable groups. Correlations in fat
metabolism, uric acid or BMI were not detected among azoospermia, subfertility and fertile control subjects.
Conclusion: No relationship was apparent between sperm recovery potential and body mass index. This
retrospective study assessed patients at a single facility; thus, future large-scale population-based longitudinalstudies are necessary.

264

●Poster Presentations

P-009

The frequency of males with sperm with low hypoosmotic swelling test scores(which prevents morphologically normal embryos from implanting) in couples
having in vitro fertilization-embryo transfer○Gabrielle Citrino, Jerome H Check, Jung K Choe, Ann DiAntonio
UMDNJ, Robert Wood Johnson Med. School at Camden, Cooper Hosp./Univ. Med. Cntr., Dept. OB/GYN, Div. Repro. Endo. &Infertility, Camden, NJ, U.S.

Introduction: The hypo-osmotic swelling (HOS) test defect is an interesting semen abnormality. If <50% of the
sperm when placed in a hypo-osmolar solution fail to show tail swelling, a defect in the functional integrity of the
sperm membrane is detected. This abnormality does not inhibit fertilization but dramatically prevents the embryosfrom implanting because of transfer of a toxic protein present from sperm to the zona pellucida which then become
incorporated in the embryo membrane causing a functional impairment of the embryo membrane, which impairs
the implantation process. Despite the fact that the knowledge that this defect markedly inhibits embryo
implantation has been known for over 25 years, and the defect is greatly obviated by intracytoplasmic sperm
injection (ICSI), this simple inexpensive test is rarely performed by most in vitro fertilization (IVF) centers. The
objective of this study was to determine how common is this abnormality in an IVF population in males withnormal semen parameters where conventional oocyte insemination rather than ICSI would generally be performed
without the knowledge of a low HOS score.Materials and methods: Over a 10 year period couples undergoing
IVF-embryo transfer (ET) were retrospectively identified where the standard semen parameters were deemed
normal (motile density >8x106/mL, morphology using strict criteria >4%, and antisperm antibodies by direct
immunobead <20%. The frequency of low HOS test scores was then determined.Results: A low HOS score wasfound in 250 of 1993 males evaluated a frequency of 12.5% in this population of males with otherwise normal
semen parameters. Conclusions: Because ICSI adds extra embryologist time, increases the expense of an alreadyexpensive procedure, and may decrease the chance of successful pregnancy and possibly increase the risk ofmono-zygotic twins it is generally not performed routinely when males have normal semen parameters. Thus IVF
centers not performing this simple inexpensive test may be choosing the wrong oocyte insemination technique andsubjecting the couple to the risks and expense of IVF with little likelihood of success when ICSI would haveworked extremely well. Hopefully, this study will encourage other IVF centers to add this simple inexpensive test
to their routine semen analysis to help prevent some couples from undergoing expensive IVF-ET without muchchance of pregnancy because of the wrong oocyte insemination technique.

P-010

Effects of several culture conditions on the primordial germ cell proliferation
1

2○Zohreh Makoolati , Mansoureh Movahedin , Mehdi Forouzandeh-Moghadam
1

3

2

Department of Anatomical sciences, Medical Sciences Faculty, Fasa University, Fasa, , Department of Anatomical sciences, Medical
Sciences Faculty, Tarbiat Modares University, Tehran, Iran , 3Department of Biotechnology, Medical Sciences Faculty, Tarbiat
Modares University, Tehran, IranObjective: The aim of this study was to compare the in vitro effects of different concentrations of retinoic acid(RA), and co-culture with STO cells on the ESCs-derived primordial germ cells (PGCs) proliferation. Materials
and Methods: CCE mouse ESCs were cultured for 1 day in order to embryoid body (EB) formation and thencultured for 4 days in the presence of 5 ng/ml BMP4 for PGC induction. In order to PGC enrichment, ESCs
derived germ cells cultured for 7 days in the presence of different doses (0-5 micromolar) of RA both in the simple
and STO co-culture systems. Expression of Mvh, PGC specific marker, was evaluated using quantitative PCR.
Data analyses were done with ANOVA and Tukey posttest. Results: The results of PCR showed higher significant
expression of Mvh in 3 micromolar RA concentrations on the top of the STO feeder layer. Conclusion:
Quantitative RT-PCR was used to estimate the level of germ cell gene expression. The results confirmed that theaddition of 3 micromolar RA concentrations on the top of the STO feeder layer has a positive role in PGC
enrichment.

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●Poster Presentations

P-011

Early development of in vitro aged eggs after intracytoplasmic sperm injection
1

1

1

1

2

○Gaku Shimoi , Masato Hayashi , Yuichi Kameyama , Ryoichi Hashizume , Ken-ichi Kudoh ,1
Masao Ito
1

Faculty of Bioindustry, Tokyo University of Agriculture, Hokkaido, Japan, 2School of Veterinary Medicine, Kitasato University,
Aomori, Japan

Introduction: Recently, rescue approaches for fertilization failure eggs (unfertilized eggs) using intracytoplasmic
sperm injection (ICSI) have been addressed (rescue ICSI). We believe that target eggs used for rescue ICSI havealready aged by the time of reinsemination and are therefore essentially different from fresh unfertilized eggs. This
study examined the development of embryos derived from the in vitro aged eggs after ICSI. Material and Methods:
BDF1 mice were used for this experiment. Superovulated eggs were cultured for 6~24 h in vitro for aging. Some ofthese aged eggs were stained by immunofluorescence, and the spindles and chromosome alignments wereobserved. For the other eggs, we performed ICSI, and fertilized eggs were cultured for 96 h in vitro. To evaluate
the quality of ICSI embryos, we performed chromosome analysis at the 8-cell stage. Additionally, we analyzed theratio of inner cell mass cells and trophectoderm cells in the blastomere by evaluating Oct4 and Cdx2 positive cells
at the blastocyst stage. Results: In 6-, 12-, 18-, and 24-h aged group, respectively, 85.9%, 57.1%, 26.9%, and 6.4%of eggs had morphologically normal spindle and chromosome alignment. The rates of these eggs in 12-h or more
aged group were significantly lower than that in the fresh group (91.0%). The fertilization rates after ICSI were
90.2% and 74.5% in 6- and 12-h aged group and 88.7% in the fresh group, and the developmental (fertilized eggs
to blastocysts) rates of these eggs were 78.4%, 25.0%, and 80.2%, respectively. The developmental rate wassignificantly lower for the 12-h aged group than for the other groups. At the 8-cell stage, the numerical aberrationof chromosome was detected in 65.5% of all observed metaphase plates in the 12-h aged group, which wassignificantly higher than that in the fresh group (29.3%). The number of Oct4 and Cdx2 positive cells in theblastocyst was 4.3 and 2.8, respectively, in the 12-h aged group, and 5.5 and 2.1, respectively, in the fresh group.
The number of Oct4 and Cdx2 positive cells in 12-h aged group was significantly lower than that in the freshgroup; however, there was no difference in the Oct4/Cdx2 ratio. Conclusions: In vitro aging caused theabnormality of cleavage apparatus in eggs and this abnormality lead to numerical aberration of chromosome andreduction of blastomere in embryos. This study suggested that the rescue ICSI should be performed for unfertilized
eggs until 6h after the first insemination.

P-012

One step closer to the development of a rapid bioassay to determine if adequate
luteal phase progesterone supplementation is provided during in vitro fertilizationembryo transfer cycles
1

1

2

1

○Ann DiAntonio , Jerome H Check , Maya D Srivastava , Rachael Cohen , Ebony Dix

1

1

1UMDNJ, Robert Wood Johnson Med. School at Camden, Cooper Hosp./Univ. Med. Cntr., Dept. OB/GYN, Div. Repro. Endo. &Infertility, Camden, NJ, 2Department of Medicine, Division of Allergy and Immunology, SUNY at Buffalo, Buffalo, NY, U.S.

Introduction: One of the functions of progesterone is to interact with a progesterone receptor that develops on
gamma delta T-cells when exposed to an allogeneic stimulus. This 34 kDa protein suppresses natural killer cellfunction by stabilizing perforin granules and causes a shift from thymic (T) helper (H) 1 cells to TH2 cells, thus
shifting the emphasis from the cellular to the humoral immune system. This protein is called the progesterone
induced blocking factor (PIBF). To date this protein has only been measured by an immunocytochemistry
technique which is not a practical assay for rapid turn-around for results. The PIBF has recently been synthesized
by recombinant DNA technology thus opening the possibility of the development of an enzyme linked
immunosorbent assay (ELISA). However for an ELISA to be developed it is first necessary to determine if PIBF isa soluable protein. Materials and methods: Serum samples were blindly assessed for the presence of PIBF using anovel sandwich ELISA we developed with full length (amino acid 1 to 757) recombinant human PIBF, rabbit
polyclonal antibody to amino acid 1-300, and affinity purified goat polyclonal antibody to the internal region of
PIBF conjugated to horseradish peroxidase. Color was developed using tetramethylbenzidine substrate stopped by
2N H2SO4 and read at 450 lambda. Bovine serum albumin served as a negative control. Results: Soluble PIBF was
detected in several samples taken from pregnant women, with highest levels in pregnant patients at over 10
microgram/ml concentration. There were significant inter-individual differences. Conclusions: PIBF, does indeed
also exist in a soluble form and can be detected in serum. With further refinement of our ELISA we aim to improvesensitivity and specificity, allowing for the simple measurement of PIBF on a rapid, high throughput basis, making
it a more practical test to determine with a bioassay if the amount of progesterone supplementation is sufficient or
not. Hopefully once a sensitive and specific PIBF ELISA is developed levels can be determined during the luteal
phase that define successful vs. non-successful embryo transfers.

266

●Poster Presentations

P-013

Effect of ovarian induction on the ultrastructure of corpus luteum during luteal
phase at implantation period
1

2

3

4

○Mandana Beigi Boroujeni , Nasim Beigi Boroujeni , Mojdeh Salehnia , Elahe Marandi ,4
5
Sadegh Rezapour , Masoud Beigi Boroujeni
1

Department of Anatomy, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran, 2Department of clinicalscience, School of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran., 3Department of Anatomy, School of
Medicine, Tarbiat Modares University, Tehran, Iran, 4School of Medicine, Lorestan University of Medical Sciences, Khorramabad,
Iran, 5Department of Biology, School of Basic Science, Shahrekord University, Shahrekord, IranIntroduction: During ovarian induction for IVF cycle, a large number of follicles develop also some ultrastructuralchanges occurred in corpus luteum that affect on pregnancy rate. The aim of this study was to evaluate
ultrastructural changes of mouse corpus luteum after ovarian induction using hMG and hCG during luteal phase at
implantation period. Material and Methods: 20 females NMRI mice 6-8 weeks old were divided into two groups:
Control group; the mice were rendered pseudopregnant and hyperstimulated group; the mice were renderedpseudopregnant after the ovarian induction. The samples were obtained from the ovary in each group at he sametime during luteal phase at implantation period. Ultrastructural changes were assessed using electron microscopy
study. Results: Our results showed that some identifiable changes were seen in structure and ultrastructure corpus
luteum in hyperstimulated group. These changes included enhancement of the apoptosis, vacuole formation andintercellular space, whereas the angiogenesis was decreased. The findings indicated decrease in organelle
condensation in cytoplasm of hyperstimulated group such as mitochondria and polyribosome, furthermore,choromatin condensation of nuclei were observed in some cell. Conclusions: Briefly, the ovarian induction usinghMG & hMG result in some ultrustuctural changes on the corpus luteum at implantation period which could be
affected on the pregnancy rate.

P-014

Autophagy in the ovarian granulosa cells○Eri Ishida, Hirohiko Tani, Miwa Shimizu, Akiyoshi Urano, Takakazu Saito, Hidekazu Saito
National Center for Child Health and Development, Department of Women's Health, Division of Reproductive Medicine and Infertility,
Tokyo, Japan

Object: Autophagy is the intracellular clearance system which breaks proteins in the cell to make a vesicle named
autophagosome. It is well known that the autophagy abundantly appears in the starvation status and the early stageof mammalian embryos just after fertilization. The cumulus cells are differentiated from the mural granulosa cells
during follicle development especially just before ovulation. The cumulus cells are associated with thetransportation of nutrition to an oocyte and the control of the meiosis of an oocyte. The association of autophagy inthe mural and cumulus granulosa cells with the maturation and quality of an oocyte has not been reported. In thisstudy, we examined the relation between the expression of autophagy and the oocyte quality using LC3 which isone of the markers for autophagosome. Method: The mural and cumulus granulosa cells was obtained from thepatients for assisted reproductive technology treatment. The oocyte quality was divided by the morphology of
cumulus oocyte complex (COC); mature, immature and dysmature. Autophagy was detected by the
immunofluorescence staining of the cumulus cells of the COC and the mural granulosa cells. Apoptosis in the
granulosa cells was also detected by the nuclear fragmentation stained with Hoechst 33342. The stain positive cells
rate was calculated in the both staining methods. Results: The rates of autophagy in cumulus and mural granulose
cells were 45.2%, 33.3%, 46.8% and 6.38% (p<0.01) (mature, immature, dysmature and mural granulosa cells
respectively). The rate of apoptosis in cumulus and mural granulose cells were 1.01%, 0.92%, 3.44% and 1.11%(p<0.01) (mature, immature, dysmature and mural granulosa cells respectively). Conclusions: In autophagy,
cumulus cells groups were higher than that of mural granulose cells. There was no difference among the cumulus
COC groups. Meanwhile the dysmature group indicated the highest incidence in apoptosis. Thus autophagy is not
related with the quality of COC.

267

●Poster Presentations

P-015

The promoter-1031(T/C) polymorphism in tumor necrosis factor-α associatedwith polycystic ovary syndrome1

12○Kwang-Hyun Baek , Ji-Hyun Yun , Jin-Woo Choi , Hyo Young Jeoung

3

1

Department of Biomedical Science, CHA Stem Cell Institute, CHA University, CHA General Hospital, Seoul, Korea, 2St. Paul's
School, Concord, NH, USA, 3Department of Obstetrics and Gynecology, CL Hospital, Gwangju, Korea

A tumor necrosis factor-α(TNF-α) is a multifunctional proinflammation cytokine that has been considered as oneof pathogenesis for various immune-related diseases. The promoter-1031(T/C) polymorphism in a TNF-αgene
was reported that it plays an part in reproduction-related diseases. And polycystic ovary syndrome (PCOS) is acommon gynecological disease of women in reproductive age women. Here, we performed a comparative study of-1031(T/C) polymorphism of TNF-αgene with PCOS in a Korean population. The -1031(T/C) polymorphism of
TNF-αgene was analyzed by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) ina total of 217 PCOS patients and 144 matched female controls of healthy women. The frequency of T/T, T/C, andC/C genotypes of -1031(T/C) polymorphism in TNF-αgene showed different proportion (p-value = 0.0003, odd
ratio (OR) = 2.53). In addition, the frequencies of co-dominant (p-value = 0.0003, OR = 2.81), and dominant
alleles (p-value = 0.0001, OR = 2.81) showed association between PCOS and control group. This is the first studyin PCOS with -1031(T/C) polymorphism in TNF-αgene. And we concluded that -1031(T/C) polymorphism in
TNF-αgene is associated with PCOS in a Korean population. Therefore, we expect that it may be considered as aclinical biomarker to diagnose for PCOS, and is helpful in understanding the etiology for the pathogenesis of
PCOS.

P-016

Association between INS-VNTR polymorphism and polycystic ovary syndrome in a
Korean population
1

1

2

2

○Bum-Chae Choi , Sang Jin Song , Ji-Hyun Yun , Bon-Hee Gu , Kwang-Hyun Baek

2

1

Department of Obstetrics and Gynecology, CL Hospital, Gwangju, Korea, 2Department of Biomedical Science, CHA Stem Cell
Institute, CHA University, CHA General Hospital, Seoul, Korea

Polycystic ovary syndrome (PCOS) is a common disorder of the women in reproductive ages. But its etiology is
not fully understood yet. Variability in the number of tandem repeats of the insulin gene (INS-VNTR) is known toassociate with PCOS, and it is associated with an increased risk of diabetes mellitus and other cardiovasculardiseases. The aim of our study was to analyze an association between the -23/Hph I polymorphism of INS-VNTR
and PCOS in a Korean population. A total of 218 PCOS patients and 141 controls were analyzed by restriction
fragment length polymorphism (RFLP) method for the INS-VNTR polymorphism. Statistical analysis ofgenotyping results were performed using HapAnalyzer. X2 test and logistic regression were utilized to analyze theassociation between two groups. A p-value under 0.05 was considered statistically significant. The frequencies of
A/A and A/T genotypes indicated a similar change between PCOS patients and controls. In addition, no association
was found between PCOS and control subjects (p-value = 0.0544, odd ratio (OR) = 1.69). Our present data
demonstrated that INS-VNTR polymorphism is not associated with PCOS in Korean women. Thus, it is suggested
that INS-VNTR polymorphism is not a key factor in the etiology and the pathogenesis of PCOS in a Korean
population.

268

●Poster Presentations

P-017

Application of non-elective blastocyst culture
Mengying Gao, ○Yanping Ma, Yonggang Li, Ze Wu , Lian Deng, Bo Deng
The Department of Reproduction and Genetics, Yunnan first people hospital, Kunming,

P.R. China

This article summarized 387 non-elective blastocyst culture and cultivation of blastocyst transfer for patients andpregnancy from February to September, 2010. We analyzed and summarized the cultivation of blastocyst transfer
and pregnancy with the 387 patients. Method: We extended the cleavage stage of embryos to the blastocyst stage invitro to observe blastocyst formation rate and quality, and analyzed the relationship between embryo quality of thecleavage stage and blastocyst formation and the impact of blastocyst formation to clinical pregnancy. The results
showed that a total of 3513 embryos of 387 patients had been cultured blastocysts and 1489 blastocyst had been
got. The blastocyst formation rate was 42.35%. I-II stage embryo blastocyst formation rate was significantly higher
than that of grade III-IVembryos (P<0.01); Clinical pregnancy rate of blastocysts was significantly statisticallysignificant at 5,6,7 days after transplantation (P<0.01). The incidence of ectopic pregnancy of blastocyst frozen
transplantation was significantly lower than that of cleavage stage embryo transfer (P<0.01). Conclusion: 1. The
rate of high quality embryonic blastocysts formation was significantly higher than low quality embryos. 2. The
formation of the pregnancy rate was higher at 5 days after blastocysts transplantation, which is low at 6,7 days. 3.
Compared with embryo transfer on day 2, blastocyst transfer was more consistent with the natural phenomenon.
Blastocyst transfer could improve the embryo implantation rate in IVF-ET and the clinical pregnancy rate andreduce the number of embryo transfer.

P-018

Oocyte activation with Ca ionophore A23187 and roscovitine for ovulated mouse
oocytes to produce haploid parthenogenones○Yuya Yano, Yuri Yamamoto, Yu Tanaka, Kenji Hinokio, Akira Kuwahara, Minoru Irahara
Department of Obstetrics and Gynecology, Institute of Health Biosciences, University of Tokushima Graduate School, Tokushima,Japan

ICSI is a essential for male factor infertility. However, total fertilization failure was observed in 2-3% of ICSI
cycles and disorder of oocyte activation is assumed to be a main cause of this unfavorable phenomenon. We
previously reported a new oocyte activation technique using a combination of Ca ionophore A23187 andpuromycin. Puromycin, a one of protein synthesis inhibitor, showed a positive effect for oocyte activation during
mouse and human oocyte, however, there is a concern of safty to use puromysin for early developmental stage ofoocyte and embryo. In this study, we evaluated the efficacy of a new combination of Ca ionophore A23187 androscovitine, which is potentselective reversible inhibitor for MPF for the oocyte activation. Retrived mice oocytes
were treated with a combination of A23187 and roscovitine (roscovitine group) or using a combination of A23187
and puromycin (puromycin group) or with A23187 (control group). Activated oocytes with 1PN2PB were observedin 88.3% in roscovitine group, 94.4% in puromycin group and 2.2% in control group. (P<0.01: control vsroscovitine or puromycin). The DNA content of activated oocytes in the roscovitine group was virtually a half ofthat of MII oocytes (47.8±10.7%). Most of activated oocytes in roscovitine group showed haploid chromosome
number (96.7%). These results indicate the efficacy and the safty of oocyte activation using a combination of
A23187 and roscovitine.

269

●Poster Presentations

P-019

The Availability of Recombinant FSH (recFSH) in Minimum Ovarian Stimulation by
the Deference of the Anti-Müllerian Hormone (AMH) Level
1

1

1

1

2

○Yuki Nagase , Miki Ikegami , Maiko Yoshioka , Tomijirou Nishihara , Mitsuru Usui ,1Toshiki Matsuura
1

Kaba Clinic Reproduction Center, Hamamatsu, Japan, 2Kyoritsu Juzen Hospital, Hamamatsu, JapanBACK GROUND: Anti-Müllerian hormone (AMH) is increasingly used to quantify ovarian reserve, but it has not
yet realized its full clinical potential in assisted reproduction technology. We usually use minimum ovarian
stimulation by only clomifene citrate (CC), in cases, by CC with recFSH. We investigated the availability ofrecFSH in minimum ovarian stimulation by various AMH level patients. MATERIALS AND METHODS: We
investigated 238 IVF cycles (mean patient age of 36.3) from August 2010 to March 2011. Patients were induced
minimum ovarian stimulation by only CC or CC with recFSH. We cultured embryo and cryopreserved at
expanding or full expanded blastocyst. Serum AMH concentrations were divided into seven groups ( <5 (a) , 5<
<10 (b) , 10< <15 (c) , 15< <20 (d) , 20< <25 (e) , 25< <30 (f) , 30< (g) pmol/L ). We investigated that we obtained
the number of eggs by an IVF cycle and the rate of blastocyst cryopreservation (BC). RESULT: We obtained morenumber of eggs with recFSH than only CC over 5 pmol/L AMH level. (only CC a: 1.2, b: 1.4, c: 1.2, d: 1.6, e: 1.9
f: 2.0, g: 1.9 eggs/IVF cycle, with recFSH a: 1.1, b: 2.6, c: 4.1, d: 4.7 e, 4.1 f: 4.0, g: 8.3 eggs/IVF cycle.) The rateof BC higher with recFSH than only CC less 10 pmol/L AMH level. The highest rate in 25<<30 pmol/L AMH
level in both group. (only CC a: 13.3, b: 21.4, c: 50.0, d: 40.0, e: 33.3 f: 100, g: 33.3 %, with recFSH a: 37.5, b:
42.9, c: 34.4, d: 38.5 e, 50.0 f: 87.5, g: 35.6 %.) CONCLUSION: This study demonstrated that minimum ovarian
stimulation by CC with recFSH effective to improvement the quality of embryo for low AMH level patients.

P-020

Results of 1027 office-based diagnostic hysteroscopies before IVF and evaluation
the pregnancy and take home baby percentages between patients with normal andabnormal uterine findings.
Selcen Bahadir, ○Mujdegul. Z Karaca, Sertac Batioglu
Zekai Tahir Burak Women's Health Education and Research Hospital, Ankara, Turkey.

Objective: To determine the findings of office hysteroscopy in an in vitro fertilization (IVF) population, also
evaluate the pregnancy and take home baby percentages between patients with normal and abnormal uterine
findings. Moreover, we compare the cost-effectivines of office hysteroscopy among other procedures. Methods:
Between May 2007 and May 2008, hysteroscopic findings in 1027 patients admitted to our IVF Unit were analyzed
retrospectively. Patients were divided into two groups, as normal and abnormal uterine findings. We examined
their record for IVF outcome. The pregnancy and take-home baby percentages were determined between thegroups. Pearsons chi-square test was used for categorical variable. p<0.05 were considered statistically significant.
Results: All hysteroscopic parameters were considered normal 792 cases (77.1%); in 235 cases (22.9%) various
pathological conditions were found. Endometrial pathology were found more frequently in older age women(p<,000). We found statistical differences for pregnancy percentages between groups with no pathology and withpathology (p<,003). Also there was a statistical differences for take-home baby percentages between these groups.
The cost of all office hysteroscopy was analyzed and compared with the other procedures. Conclusions: There wasa trend toward better pregnancy and take-home baby percentages in women with normal hysteroscopic findings.
Because of this routine uterine cavity evaluation should be perfomed in all patients. But for first line examination,
non invasive and cheap procedures should be recommended.

270

●Poster Presentations

P-021

Serum levels of macrophage inhibitory cytokine 1 (MIC 1) as a predictor of
miscarriage in early pregnancy of women treated for repeated reproductive failures○Daniela. N Baltadzhieva, Kalinka. L Penkova, Pepa. A Angelova, Meglena. M Metodieva
Center for Reproductive Health "Nadejda", Sofia, Bulgaria

Imbalances in the intrauterine cytokine milieu around the time of implantation may play a causative role in earlypregnancy failure. Cytokines appear to be an important component of a communication network at the
feto-maternal interface. Experimental evidence suggests that MIC-1 has immunomodulatory activity and acts tomaintain pregnancy by suppressing the production of proinflammatory cytokines. Studies of serum MIC-1 levels inseveral cohorts of patients have revealed potential clinical use in the monitoring of pregnancy. The aim of ourstudy was to evaluate the prognostic value of serum MIC-1 levels in pregnant women treated for repeated
reproductive failures. In the study, 59 women treated for repeated reproductive failures (15 women with RSA and
44 women with RIF) were included at positive blood pregnancy test. Serum samples were collected from thewomen between 4-16 weeks of pregnancy and MIC-1 concentrations were measured using a sensitive
enzyme-linked immunosorbent assay (BioVendo, CzR). Serum levels of MIC-1 in 5th week of gestation weresignificantly higher (p=0.033) in the group of women pregnant by ART procedure in comparison with the group of
spontaneously pregnant women. No difference in MIC 1 levels was observed between the groups in latergestational weeks. Eight women (13.56%), 2 from the RSA group and 6 from RIF group, miscarried in 9-10 weekof gestation and their MIC 1 concentrations were significantly lower (p=0.045) than those with ongoing
pregnancies. Low MIC-1 serum levels were detected before miscarriage in the pregnant women treated for
repeated reproductive failures, suggesting possible predictive and causative role of this cytokine and that theapplication of therapeutic agents that would lead to up-regulation of MIC-1 prevent miscarriage.

P-022

A drop in serum estradiol the day after human chorionic gonadotropin (hCG) shot
does not adversely affect pregnancy rates per embryo transfer in women with very
decreased oocyte reserve○Ann DiAntonio, Jung K Choe, Jerome H Check
UMDNJ, Robert Wood Johnson Med. School at Camden, Cooper Hosp./Univ. Med. Cntr., Dept. OB/GYN, Div. Repro. Endo. &Infertility, Camden, New Jersey, U.S.

Introduction: The majority of hyperstimulated women having in vitro fertilization-embryo transfer (IVF-ET) show
a rise in serum estradiol (E2) the day after hCG. However, women with markedly diminished oocyte reserve withusually no more than 1-3 follicles may follow the course more akin to normal ovulatory cycles when the serum E2
drops after the luteinizing hormone (LH) surge as progesterone (P) rises. The objective of the present study was todetermine if such a drop in this special group of patients has any negative impact on the pregnancy rate (PR) or
implantation rate following ET. Materials and methods: Over a 10 year period women undergoing IVF-ET with aday 3 serum follicle stimulating hormone (FSH) >15 mIU/mL were identified. Only the first cycle of IVF-ET was
evaluated for serum E2 and P and E2/P ratio on the day of and the day after hCG injection. They were divided into
2 groups: those with an increase vs. those women with a decrease in serum E2 the day after hCG injection. P level
and E2/P ratio were also compared not only between these 2 groups but in those who conceived vs. those who did
not. Comparisons were made with chi-square analysis. Results: There were 403 IVF-ET retrievals selected: 325
with E2 rising (grp 1) and 78 with E2 dropping (grp 2). Thus 19.3% showed a drop in E2. The clinical and live
delivered PRs/transfer were similar in grp 1 and grp 2, 31.1% (61/196) and 28.1% vs. 38.6% (17/44) and 22.7%(p=NS, chi-square). Implantation rates were also similar, 22.1% (70/317) vs. 28.0% (21/75). There was nodifference in the level of E2 on day of hCG, 490pg/mL grp 1 vs. 478 pg/mL grp 2. Comparing E2/P ratio inpregnant vs. non-pregnant women grp 1 vs. grp 2 provided no new insights. For grp 1, 49 released the oocyte(15.0%) before the retrieval vs. 14 (17.9%) in grp 2 and no eggs were retrieved in 39 (12.0%) grp 1 vs. 13 (16.7%)
grp 2 women (p=NS). Conclusions: A decrease in serum E2 the day after hCG shot in women with diminishedoocyte reserve has no adverse consequences for IVF-ET.

271

●Poster Presentations

P-023

Analysis of the limitations for the numbers of attempts of infertility treatment using
ART.
○Hiroaki Shibahara, Tatsuya Suzuki, Kenro Chikazawa, Tomoe Ikeda, Yuki Hirano, Mitsuaki Suzuki
Department of Obstetrics and Gynecology, School of Medicine, Jichi Medical University, Tochigi, Japan

Purpose: The successful treatments using ART, including IVF-ET, ICSI-ET and frozen-thawed ET, have offeredsatisfactory results for a number of infertile couples. However, there still exist refractory infertile couples even they
are treated by modern ART. The aim of the present study is to analyze the limitations for the numbers of attempts
by ART, the final stage of infertility treatment, especially in such patients. Method: Between Jan. 2006 and Dec.
2009, 157 infertile couples were treated by ART only in our institute and the final results of their treatments wereobtained. Patients who were not yet completed their treatments or those who had experienced the ART treatments
in other clinics were excluded. In all, 247 fresh embryo transfers and 163 frozen-thawed embryo transfers were
included. As for blastocyst transfers, the results in 115 treatment cycles from 45 couples were analyzed. Results:
The pregnancy rates (PR) per cycle and that per patient were 37.2% and 58.6%, respectively. The multiple PR was
5.4%, including only twin pregnancies. In all, the cumulative PR reached plateau (98.9%, 95.7%) when thenumber of oocyte picking-up and that of embryo transfer carried out were 3 cycles and 4 cycles, respectively. The
approximate limitation of the total numbers of embryos for achieving pregnancy was designated as 7. As forblastocyt transfers, the cumulative PR reached plateau (90.3%) when the number of embryos transferred was 3.
The approximate limitation of the total numbers of blastocysts for achieving pregnancy was designated as 6.
Conclusions: These findings should be informed for the decision making before the patients start to be treated by
ART.

P-024

The predictive value of AMH for ovary reserve and response during IVF-ET○Xiaomei Zhang
The reproductive medicine center of Subei People's hospital, China

Introduction: There is an urgent need for a reliable and early marker for the detection of a declining number offollicles, and prediction of spontaneous pregnancy potential and assisted reproduction technology outcomes.
Recent studies shown that Anti-Mullerian hormone (AMH) maybe such a promising and reliable marker. We
therefore carried out a prospective trial to study the predictive value of AMH for ovary reserve and response during
IVF-ET. Material & Methods: (1) The level of AMH protein secretion in serum was examined by ELISA in 228
cases of infertile woman, the level of AMH protein in follicular fluid and the AMH mRNA relative concentrationin granulosa cells were examined by ELISA and RT-real-time-PCR in 71 cases who received IVF-ET treatment.
Results: (1) A positive correlation was found between serum AMH and AFC (p<0.01) in 228 cases of infertile
women, and it correlated negatively with Age, Age at Menarche and Years Since Menarche (p<0.01, p<0.01,
p<0.01), serum AMH level and AFC correlated each other and declined significantly with age. (2) There is no
significant difference between the bAMH with the AMH protein level on day 5 after Gn injection and on oocyte
retrieval day (p>0.05) in 71 cases of IVF-ET treatment patients. (3) A positive correlation was found between
serum AMH with FF AMH and relative concentration of AMH mRNA in GC (p<0.01,p<0.01) in 71 cases of
IVF-ET treatment patients, there are significant differences comparing serum, FF AMH level and GC AMH
mRNA relative concentration with the ovary response (p<0.05). (4) A positive relationship was found between
bAMH and the number of oocytes retrieved (p<0.01), and there are positive correlations between the AMH mRNAlevel with the number of fertilized oocytes and the number of cleavage (p<0.01,p<0.01) (5). The basic AMH levelin 12 OHSS patients is significantly higher than the 12 ovary poor response cases (p<0.01). Conclusions: There is
obvious relationship between serum AMH level with ovary reserve and response. The serum AMH protein level isof more clinical value to predict the ovary reserve and response, and may be applied in clinic as a molecule marker.

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●Poster Presentations

P-025

Whether Day 3 FSH/LH ratios is a good predictor of IVF prognosis in GnRH
antagonist protocol?
1

2

1

1

1

○K.H. Lee , J.D. Cho , H.G. Sun , S.K. Kim , J.H. Lee , Ilhae Park

1

1

Mamapapa&baby OB&GY, 2Ellemedi Infertility Clinic, Changwon, Gyoung Nam, repubilic of Korea

Introduction: As most of us in this field know, Day 3 FSH is a simple and easily-available ovarian reserve marker
and that, its value is limited when FSH is significantly elevated. We also know that Day 3 LH also has limited
value in ovarian reserve marker. Some previous studies have shown FSH/LH ratios could be used as ovarian
reserve marker. The problem was that these studies were limited by small sample sizes and almost all studies were
performed in GnRH agonist protocol and not in GnRH antagonist protocol. Objective: The purpose of this study
was to examine whether day 3 FSH/LH ratios could predict IVF outcome in GnRH antagonist cycles. Design: A
retrospective cohort study. Materials and methods: We retrospectively reviewed 228 patients undergoing IVF using
the GnRH antagonist protocol from January 2004 to December 2008. Inclusion criteria included age < 40, day 3
FSH < 10 IU/L. Day 3 FSH/LH level of included patients were checked within 3 months of IVF start. The patientswere divided into 3 groups. The groups were based upon Day 3 serum FSH/LH ratios. Group I consisted of 117
patients with serum FSH/LH < 2, Group II consisted of 64 patients with 2< FSH/LH <3 and Group III consisted of
47 patients with FSH/LH > 3. We compared characteristics and cycle outcomes of patients. Results: The
characteristics of patients by age, parity and cause of infertility were similar among the 3 groups. The number ofoocytes (Group I: 14.5, Group II: 12.5, Group III: 9.46) and the number of 2PN ( Group I: 10.5, Group II: 8.7,
Group III: 6.18) were significantly decreased when day 3 FSH/LH ratios increased. Ongoing pregnancy rates
significantly decreased as FSH/LH ratio increased. [Group I: 47.8%(56/117), Group II: 35.9%(23/64), Group III:19.1%(9/47), P<0.01] Conclusions: We observed that increased FSH/LH ratios were associated with poor IVF
outcomes in GnRH antagonist cycles. Moreover, as FSH/LH ratios increased, the number of oocytes, the numberof 2PN and ongoing pregnancy rates decreased. Therefore, we suggest that Day 3 FSH/LH ratios can be used as
simple and easily-available ovarian reserve marker and a good predictor of IVF prognosis.

P-026

Luteal blood flow in patients undergoing GnRH agonist long protocol○Kumiko Mizumoto, Akihisa Takasaki, Maki Okada, Yuuko Sakaguchi, Katsunori Shimamura,
Hitoshi Morioka
Saiseikai Shimonoseki Hospital, Yamaguchi, Japan

Background: Blood flow in the corpus luteum (CL) is closely related to luteal function. It is unclear how
lutealblood flow is regulated. Standardized ovarian-stimulation protocol with a gonadotropin-releasing hormone
agonist(GnRHa long protocol) causes luteal phase defect because it drastically suppresses serum LH levels.
Examiningluteal blood flow in the patient undergoing GnRHa long protocol may be useful to know whether luteal
bloodflow is regulated by LH. Methods: Twenty-four infertile women undergoing GnRHa long protocol weredivided into 3 groups dependenton luteal supports; 9 women were given ethinylestradiol plus norgestrel (Planovar)
orally throughout the lutealphase (control group); 8 women were given HCG 2,000 IU on days 2 and 4 day after
ovulation induction inaddition to Planovar (HCG group); 7 women were given vitamin E (600 mg/day) orally
throughout the luteal phasein addition to Planovar (vitamin E group). Blood flow impedance was measured in each
CL during the mid-lutealphase by transvaginal color-pulsed-Doppler-ultrasonography and was expressed as a
CL-resistance index (CL-RI). Results: Serum LH levels were remarkably suppressed in all the groups. CL-RI in thecontrol group was more thanthe cutoff value (0.51), and only 2 out of 9 women had CL-RI values < 0.51.
Treatments with HCG or vitamin Esignificantly improved the CL-RI to less than 0.51. Seven of the 8 women in the
HCG group and all of the womenin the vitamin E group had CL-RI < 0.51. Conclusion: Patients undergoing
GnRHa long protocol had high luteal blood flow impedance with very lowserum LH levels. HCG administration
improved luteal blood flow impedance. This suggests that luteal blood flow isregulated by LH.

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●Poster Presentations

P-027

Comparison of the effect of GnRHa long and short protocol in older women during
IVF/ICSI○Xun Zeng, Shangwei Li, Lang Qing, Xiaohong Li, Han Hu
Reproductive Medical Center of West China 2nd Hospital,Sichuan,China

Objective: This retrospective study aimed to compare the effect of GnRHa long and short protocol in women aged
35 and above after IVF/ICSI attempts. Methods: A total of 401 fresh IVF/ICSI cycles were followed in my study.
In women aged 35-39 years group, 233 cycles were included in the long protocol and 105 cycles in the shortprotocol. In another age group, women aged 40 years and above, 14 and 49 cycles were included in the long andshort protocol, respectively. Results: In women aged 35-39 years group, there were significant differences in thedays of Gn using (9.75±2.03 vs 8.26±1.76, P<0.05) and the amount of Gn (31.57±9.38 vs 26.48±7.64, P<0.05) in
GnRHa long and short protocol respectively. The EMT on the dayHCG (10.00±2.26mm, P<0.05) in long protocol
were thicker than that in short one (9.20±1.90mm, P<0.05). The number of oocytes retrieved (7.46±4.77 vs5.2±3.96), cleavage (4.65±4.04 vs 3.56±3.07), excellent embryos (3.36±2.12 vs 2.31±1.99) and the rate ofexcellent embryos (69.07% vs 60.19%) in the long and short protocol were significant difference (P<0.05), as wellas the implantation rate (21.5% vs 13.0%, P<0.05) and the pregnancy rate (39.7% vs 26.8%, P<0.05). In anotherage group,the days of Gn using (10.60±1.34 vs 7.74±1.82) and the amount of Gn (37.50±7.49 vs 26.25±7.59) inlong and short protocol respectively were significantly different (P<0.05). There were significant difference in thenumber of oocytes retrieved (6.91±6.75 vs 3.80±3.14, P<0.05), excellent embryos (2.43±1.16 vs 1.47±1.30,
P<0.05) and the rate of excellent embryos (68.56% vs 53.62%, P<0.05). But there were no significant difference in
implantation rate and pregnancy rate. Conclusion: women aged 35 years and above who received long protocol
needed more ampoules of Gn and longer period of treatment. More oocytes, a greater rate of high quality embryos
and a higher clinical pregnancy rate can be obtained in GnRHa long protorol.

P-028

Small amount of testosterone administration improves ovarian response toovulation induction in poor responders
1

1

1

1

2

○Kohzo Aisaka , Haruko Hiraike , Hiroe Hyodo , Seiichiro Obata , Osamu Hiraike , Hironobu
2
Hyodo
1

Department of OB/GYN, Hamada Hospital, Tokyo, 2Department of OB/GYN, University of Tokyo, Japan

Objective: It is well known that there are some patients who cannot respond properly by the exogenous
administration of gonadotropin preparations. It is also reported that plasma estrogen can modify the activities ofgonadotropin receptors. We have already reported that exogenous administration of estrogen is effective toimprove the response of gonadotropin administration in these patients, however there are still some patients who
cannot be treated by this method. Present study was performed to elucidate whether exogenous administration of
estrogen with a small amount of testosterone was effective to improve the response of gonadotropin administrationin the patients of severe poor responders. Method: Six patients who had resistance to gonadotropin therapy, andcould not succeed by the administration of the exogenous estrogen therapy were subjected (35.4+/-1.9 years old).
All of them had experiences of previous treatment of hMG (up to 600/day), and could not respond to the treatment.
Then, exogenous estrogen (conjugated equine estrogen: 2.5 mg/day) and testosterone cream (trans-dermal,
0.6mg/day) was administered for 2 weeks with Gn-RH agonist. After that, hMG preparation of 150IU/day wasadministered to the subjects with estrogen, testosterone and Gn-RH agonist in gradual increasing method up to
600/day, and the follicular growths were observed by the transvaginal ultrasonic scanner. Then, 10000iu of hCG
was injected when the matured follicles (diameter over 18mm) were observed, and the proper luteal support(progesterone vaginal tablets, 60 mg/day) was also done. Results: The ovulations were observed in four of six
patients, and one could conceive with this method. Conclusion: The exogenous testosterone administration withestrogen and Gn-RH agonist is effective to the ovulation induction in the patients of severe poor responders. It wassuggested that the exogenous testosterone and estrogen might have some effect to improve the sensitivity of the
gonadotropin receptors in the ovaries.

274

●Poster Presentations

P-029

rFSH combined with uhMG had significantly increased pregnancy compared withrFSH alone undergoing ovarian stimulation following a long protocol in IVF
1

2

1

1

1

○K.H. Lee , J.D. Cho , H.G. Sun , I.H. Park , J.H. Lee , S.K. Kim

1

1

Mamapapa&baby OB&GY, 2Ellemedi Infertility Clinic, Changwon, Gyoung Nam, Repubilic of Korea

Objective: The purpose of this study was to compare the pregnancy rates resulting from ovarian stimulation withrFSH combined with uhMG or rFSH alone in women undergoing IVF cycles. Material & methods: The study
included 144 cycles (134 patients) between January 2007 and December 2007 in IVF cycles, and controlled
ovarian hyperstimulation were performed with GnRH agonist long protocol. The patients were classified in two
groups: Group A- rFSH alone for controlled ovarian hyperstimulation following a GnRH agonist long protocol (69cycles, n=63). Group B- rFSH combined with urinary hMG for controlled ovarian hyperstimulation following a
GnRH agonist long protocol (75 cycles, n=71). We compared patients age, the number of retrieved oocytes,
transferred embryos, the percentage of metaphase II oocytes and clinical pregnancy between the two groups.
Results: The age of the patients, the percentage of metaphase II oocytes, the number of transferred embryos andabortion rates were comparable in two group. The number of retrieved oocytes (17.6 ± 8.7, 12.3 ± 6.3; P<0.001)
and embryos (13.0 ± 7.1, 8.5 ± 4.9; P<0.001) were significantly higher for group A than for group B. However,pregnancy outcomes including clinical pregnancy rates (55.1%, 64.0% P<0.01), ongoing pregnancy rates (44.9%,
52.0% P< 0.01) and implantation rates (19.6%, 20.7% P<0.01) were significantly higher for group B than for group
A. Conclusions: Our data indicates that the use of rFSH combined with urinary hMG was significantly higher
clinical pregnancy rate than rFSH alone undergoing ovarian stimulation following an agonist long protocol.
Comparing our analysis of the two groups, the use of rFSH combined with urinary hMG had significantly
increased pregnancy, despite a lower number of retrieved oocytes and fertilized oocytes. The hypothesis was thatembryo quality and endometrial receptivity were significantly improved after using rFSH combined with urinary
hMG, compared with rFSH alone, probably due to the presence of LH. Therefore, LH activity may influence
treatment response and outcome in IVF cycles. Further studies were that data from a larger population are needed
to precisely estimate any difference between uhMG and rFSH.

P-030

A comparison of pregnancy outcome in women with normal oocyte reserve
according to the use of high or low dose follicle stimulating hormone (FSH)
stimulation○Gabrielle Citrino, Jerome H Check, Jung K Choe
UMDNJ, Robert Wood Johnson Med. School at Camden, Cooper Hosp./Univ. Med. Cntr., Dept. OB/GYN, Div. Repro. Endo. &Infertility, Camden, NJ, US

Introduction: The main advantage of high dosage FSH controlled ovarian hyperstimulation (COH) is a greater
embryo yield allowing better odds of having top quality embryos and more cryopreserved embryos for futuretransfers. The main advantage of low FSH stimulation is reduced expense and less risk of ovarian hyperstimulation
syndrome (OHSS). The objective was to determine if more embryos from which to choose provides higher
pregnancy rates (PRs) per transfer or could the need to cryopreserve for OHSS risk reduce the chance of pregnancy
on first transfer (fresh or frozen)? Finally, would the chance of having more embryos with high stim result in agreater chance of having an extra frozen embryo transfer resulting in a higher PR per oocyte
harvest&quot.Materials and methods: A retrospective review was done on 1st oocyte retrievals over a 10 year
period on women with normal ovarian reserve. The data were stratified according to 2 age groups (35 and 36-39)
and according to whether high dose FSH COH (187.5 IU) or low dose FSH (>187.5 IU) was used. Comparisonswere made according to the first fresh ET, the PR per 1st transfer including a frozen ET and per oocyte harvest.
Results: Fresh transfers only clinical and live delivered PR, abortion rate, and implantation rate for women 35: highdose FSH 50.2% (210/418), 41.9%, 16.7% (35/210) and 32.1% (322/1004); low FSH 51.0% (99/194), 44.8%,
12.1% (12/99) and 34.6% (149/431). First transfer (fresh or frozen) 49.2% (245/498), 41.2%, 16.3% (40/245) and
31.0% vs. 49.6% (118/238), 43.7%, 11.9% (14/118) and 33.0% (178/539). PR per harvest 64.4%, 53.0%, and17.7% vs. 57.0%, 49.4%, and 13.2%. For ages 36-39 the comparative values for fresh ET were for high FSH
stimulation 35.4% (92/260), 26.9%, 23.9% (22/99) and 19.7% (137/694) vs. 28.7% (27/94), 25.5%, 11.1% (3/27)
and 20.1%. First transfers 33.9% (99/292), 26.4%, 27.3% (27/99), and 18.6% (146/787) vs. 32.7% (34/104),
28.8%, 11.8% (4/34) and 22.2% (51/230). PR per harvest 41.8% (135/323 retrievals), 32.5%, and 22.2% (30/135)
vs. 30.5% (40/131 retrievals), 26.7% and 12.5%). Conclusions: The main advantage of high stimulation over low
stimulation from a PR standpoint is that women aged 36-39 have a 25% higher pregnancy rate per harvest.
Otherwise, the outcome between the two methods were similar. The couple must weigh reduced risk and cost withlow stim vs. one additional benefit for high stim, i.e., possibly more embryos for future transfers.

275

●Poster Presentations

P-031

The effects of laser assisted hatching on pregnancy rates○Forouzan Absalan

1,21Anatomical department, Jundishapoor university of medical sciences, Ahvaz, , 2Iranshiraz infertility and limited surgical center, Shiraz,
Iran

Background: For infertile women aged over 35 years, failure of the ZP (zona pellucida) to rupture is believed to be
associated with a decreased implantation rate in In Vitro Fertilization (IVF) or Intra Cytoplasmic Sperm Injection(ICSI). Objective: In this research Laser assisted hatching was offered to patients with advanced maternal age toevaluate a possible benefit. Materials and methods: Nine hundred thirty two cycles of IVF/ICSI in females wereanalyzed. Women included in this study were allocated in 4 groups. In group I and II, embryos were cultured andtransferred with and without LAH in women aged under 35, where as embryos of group III and IV were examinedwith and without LAH in women aged over 35. Laser manipulations were performed using a suturn Tm3 systemusing 2-3 pulses of 0.8 millisecond with 400 voltage duration. The size of the hole made in the zona was measured
to be 5-10 µm, depending on the zona thickness of each individual embryo.Results: The performance of LAH
significantly increased clinical pregnancy rates in all patients. The chemical pregnancy (50.99% and 31.61%
respectively), clinical (50% and 30.69% respectively) and multiple pregnancies (22.27%and 5.94% respectively)
significantly differ between group I and II. In the patients with advanced female age over 35 the performance of
LAH significantly increased chemical (30.12%) and clinical pregnancy (27.71%) rates compared to whom thatwithout LAH (18.96%) and (16.37%) respectively. Conclusion: Our data demonstrate in the patients who were lessthan 35 years old, multiple pregnancy rates were significantly increased compared other groups who aged over 35
years old. In addition benefit of LAH in improving pregnancy ratesafter IVF or ICSI in women of advanced age(over 35) was shown.

P-032

Correlation between semen intracellular parameters and fertilization rates
following ICSI
○Mina Sharbatoghli, Bita Ebrahimi, Mojtaba Rezazade Valojerdi
Emberyology Department, Cell Sciences Research Center, Royan Institute, ACECR, Tehran, IranIntroduction: Nowadays, using functional tests alongside with the conventional semen analysis could be effective
in foreseeing the treatment results. This study was set out to investigate how mitochondrial membrane potential,
DNA fragmentation index and apoptosis in ejaculated spermatozoa correlate with each other and also laboratory
outcomes after intracytoplasmic sperm injection (ICSI). Materials and methods: We examined ejaculated
spermatozoa from 120 patients undergoing ICSI treatment. Sperm DNA fragmentation Index was assessed bysperm chromatin dispersion test (SCA). Mitochondrial membrane potential (MMP) and incidence of apoptosis
were also evaluated by flow cytometry. Result: There were significant positive correlation between apoptosis,
MMP and DNA fragmentation index but these parameters did not show any significant correlation with laboratory
ICSI outcomes. Conclusion: Despite the existence of a relationship between intracellular factors, in this study no
relationship between these parameters with fertilization rate was observed. This result could be caused by choosing
high quality sperms according the classical factors by an embryologist in ICSI process and more research might behelpful.

276

●Poster Presentations

P-033

Whole ooplasmic transfer by direct injection method using Piezo driven system forprevention of mutated mitochondrial DNA transmission
1

1

1

1

1

○Akiko Yabuuchi , Noriko Kagawa , Kenji Ezoe , Chiemi Mori , Yuko Takayama ,
2
1
1
1
1
Masashige Kuwayama , Keiichi Kato , Fumihito Aono , Yuji Takehara , Osamu Kato
1

Advanced medical research institute of fertility, Kato Ladies Clinic, Shinjuku, Tokyo, Japan, 2Repro-support medical research center,
Shinjuku, Tokyo, Japan

Mitochondrial diseases are caused by pathogenic mutation in mitochondrial DNA (mtDNA) and treatment option
for patients having mitochondrial disease are severe. Mitochondrial DNA is transmitted maternally and in Assisted
Reproductive Technology (ART), it has been proposed that ooplasmic transfer which is the technique to exchange
mutated mtDNA to normal mtDNA by replacing oocyte cytoplasm using nuclear transfer techniques is thepotential treatment to prevent mtDNA transmission. Ooplasmic transfer has been applied by using existing method,i.e. cell membrane fusion method in animals and human. However, these reconstructed oocytes are caused mtDNA
heteroplasmy as donor karyoplast (donor nuclei with cytoplasm) including donor mtDNA is fused with recipient
cytoplasm which has mtDNA from recipient result in mixture of two different mtDNA in the oocyte. Therefore, itis necessary to invent new techniques to avoid mtDNA heteroplasmy. Here, we developed innovative technique ofooplasmic transfer in germinal vesicle (GV) oocytes which we inject GV (donor nuclei only, with no cytoplasm)
directly into oocyte using piezo drive system termed as Germinal Vesicle Injection (GVI). Bovine GV oocytes
were used as their oocyte size is similar to human GV oocytes. Donor GV was taken from oocytes after disrupting
oocyte membrane, removed cytoplasm attached around GV membrane and then directly injected to recipient
enucleated GV oocytes using Piezo drive system (Primetech Co. Ltd., Japan). Inner diameter (ID) of injection
pipette should be 30 μm adjusted outer diameter of bovine GV which has been thought that 30 μm ID of injection
pipette is invasive for the oocytes. But we improved the survival rate of oocytes after injection up to 95% (58/61)
by using 1 μm of thickness of pipette with minimum intensity of piezo pulses. In vitro maturation and in vitro
developmental rate to the blastocyst stage after fertilization of GVI oocytes were 81% (47/58) and 22% (8/47),
respectively and there were no significant differences between non treated GV oocytes (81%, 70/86 and 25%,
11/44). These results indicate that our new GVI technique is effective and non invasive method for bovine oocytes.
We hypothesize that our GVI technique will be able to avoid mtDNA heteroplasmy if all mitochondria attachedtightly around donor GV are completely removed or inactivated. More detailed experiment should be performed
before clinical trial. But we anticipate that our GVI tecnique is the potential treatment to prevent mutated mtDNA
transmission.

P-034

Elevation of the Risk of Monozygotic Twinning by Expanded Blastocyst Transfer○Kazusuke Nagoshi
Nagoshi Ladies Clinic, Okayama, Japan[Objectives] During in vitro fertilization and culture, monozygotic twinning (MZT) cannot be completely avoided
even when fertilization is performed by single embryo transfer, and the rate of MZT after in vitro fertilization ishigher than that after natural fertilization. MZT is considered to occur through the splitting of the inner cell mass
during hatching via the narrow opening of the zona pellucida. We recently encountered 6 cases where MZT
resulted from single blastocyst transfer, which was performed after manipulation to ensure complete hatching via
the zona pellucida. We report these cases with discussions about the methods of fertilization and the backgroundfor embryonic development. [Subjects] Among all cycles of embryo transfer performed in our clinic between
January 2006 and April 2010, 494 cycles involved single blastocyst transfer (95.5% of the cases), and pregnancy
resulted from 196 cycles (pregnancy rate, 39.7%). The subjects who underwent these 196 cycles were the subjectsof this study. During embryo transfer in each case, complete hatching via the zona pellucida was ensured by aphysical method. [Results] MZT occurred in 3.1% (6/196) of pregnancies. The MZT rate was 4.8% (4/84) withfresh embryo transfer and 1.8% (2/112) with vitrified-warmed embryo transfer. The MZT rate was analyzed inrelation to the fertilization method: the rate was 3.9% (4/103) with conventional in vitro fertilization(C-IVF)-derived blastocyst transfer and 2.2% (2/93) with intracytoplasmic sperm injection (ICSI)-derived
blastocyst transfer. The rate of MZT after fresh embryo transfer was 6.7% (3/45; C-IVF-derived embryo) vs.
2.6% (1/39; ICSI-derived embryo). The MZT rate after vitrified-warmed embryo transfer was 1.7% (1/58;
C-IVF-derived embryo) vs. 1.9% (1/54; ICSI-derived embryo). Further, the MZT rate was analyzed in relation to
the embryo grade: the rate was 3.3% (6/184) after transfer of high-grade embryo (at least 3BB) and 0% (0/12) after
transfer of low-grade embryo. Of the high-grade embryos whose transfer resulted in MZT, the growth stage at thetime of transfer was blastocyst in 0% embryos, expanded blastocyst in 6.9% (4/58) embryos, and hatching/hatchedblastocyst in 2.2% (2/92) embryos.

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●Poster Presentations

P-035

Evaluation of sperm chromatin integrity following IMSI or HBA for pre-ICSI sperm
selection○Satoshi Ueno, Kazuo Uchiyama, Keiichi Kato, Yuji Takehara, Osamu Kato
Kato Ladies Clinic, Tokyo, Japan

Objective: For almost two decades ICSI has been successfully used to obtain high fertilization and pregnancy rates
in couples with severe male infertility. In recent years however new sperm selection methods such as
intracytoplasmic morphologically selected sperm injection (IMSI) and hyalorunan-binding-assay (HBA) has beendeveloped to identify the most competent gametes before ICSI procedure. In this prospective study we evaluatedthe integrity of sperm chromatin following selection with the above methods. Materials and methods: Afterobtaining informed consent ejaculated sperm samples were collected from infertile patients requiring ICSI
treatment. Samples were initially prepared using discontinuous gradient centrifugation re-suspended and divided
into four groups. IMSI group (A): normal morphology sperm containing no vacuoles in the nuclear area wasselected by observation under x1000 magnification inverted microscope. HBA group (B): in Petri dish a 2μl
droplet of hyalorunan (HA) containing medium (Sperm Slow, MediCult, Denmark) was connected to anotherdroplet containing the suspended sperm. After incubation the HA-bounded sperm cells were retrieved from thejunction zone of the two droplets. ICSI control group (C): sperm with normal morphology and excellent motility
was selected by observation under x 200 magnification conventional ICSI equipment. Unselected control group(D): sperm was randomly selected from the initially prepared suspensions. Sperm chromatin integrity as the main
outcome measure was evaluated by sperm chromatin dispersion test using a commercially available kit(Halosperm, Halotech, Spain). Pair wise comparisons were made between the experimental and control groups
using chi-square test to check for statistical significance. Results: The sperm chromatin fragmentation rates were
4.5% (2/48), 2.6% (4/151), 10.9% (7/71) and 19.9% (48/289) for groups A, B, C and D, respectively. Differenceswere statistically significant in favor of HBA group compared to group C (p=0.03) and D (p<0.0001). Compared tothe two control groups the IMSI group showed a non significant trend towards lower sperm chromatin
fragmentation values. Conclusions: Our preliminary data suggests that both HBA and IMSI are effective methods
for non-invasive selection of sperm cells with high sperm chromatin integrity. Further studies are needed toevaluate the impact of these new sperm selection techniques on assisted reproduction treatment outcome.

P-036

Differential effects of urinary and recombinant chorionic gonadotropin on oxidative
stress responses in decidualizing human endometrial stromal cell.
○Takeshi Kajihara, Japarath Prechanich, Hideno Tochigi, Osamu Ishihara
Saitama Medical University, Saitama, Japan(Introduction) Human chorionic gonadotropin (hCG) is one of the earliest signals secreted by the implanting
embryo. In addition to its well-known luteotropic function in early pregnancy, hCG also acts directly on
decidualizing endometrium. Recently, we demonstrated that recombinant hCG (rhCG) prevented apoptosis in
decidualizing human endometrial stromal cells (HESCs) exposed to oxidative stress. Two hCG preparations arewidely used clinically: rhCG, produced by recombinant DNA technology, and urinay hCG (uhCG), extracted fromurine of post-menopausal women. However, an analysis of the direct effects of rhCG and uhCG on the decidual
phenotype of HESCs has not yet been done. In this study, we investigated the effects of uhCG and rhCG on themorphological and functional profiles of decidualizing HESCs. (Material & Methods) HESCs from hysterectomy
specimens were isolated and incubated with 8-bromo-cyclic adenosine monophosphate and medroxyprogesterone
acetate in the presence or absence of recombinant hCG (rhCG) or urinary hCG (uhCG). Hydrogen peroxide wasused as the source of reactive oxygen species (ROS). The amounts of PRL in culture media were examined.
Western blot was performed for protein analysis. The level of apoptosis was analyzed by cell detection ELISA Kit.
(Results) Neither rhCG nor uhCG alter the morphological appearance of the decidual HESC cultures, although
rhCG attenuated prolactin expression, a major decidual marker protein. rhCG represented protective effect on
decidualizing HESCs to oxidative cell death through enhanced expression of SOD2, a cardinal enzyme in thecellular defense against oxidative damage. rhCG signaling also selectively limits activation of the apoptotic
machinery in decidualizing HESCs by enhanced Bcl-2 expression whereas uhCG induces the expression of Fas
ligand. (Conclusion) Our results suggest that rhCG might be a preferable agent to protect the maternal decidua
against oxidative damage in pregnancy, especially at the time of implantation and beyond.

278

●Poster Presentations

P-037

Heparin prevents programmed cell death induced by oxidative stress in humandecidualized endometrial stromal cells
Takeshi Kajihara, ○Japarath Prechanich, Hideno Tochigi, Osamu Ishihara
Saitama Medical University, Saitama, Japan(Introduction) Heparin is clinically administered for the prevention of thromboembolic disorders during pregnancy,
especially for antiphospholipid antibody syndrome. A recent clinical trial demonstrated a beneficial effect ofheparin administration in the luteal phase on implantation rate as well as the live birth rate in women with repeatedimplantation failure. In addition, heparin exerted direct effects on human endometrial stromal cells (HESCs),
independently of its anticoagulant activity. However, the precise effects of heparin on decidualization process
ramains uncertain. Recently, we demonstrated that human endometrial stromal cells (HESCs) become
extraordinarily resistant to oxidative stress-indued apoptosis upon decidualization. Therefore, we hypothesized thepossible direct action of heparin on HESCs decidualization, that might induce the expression of genes participatingin resistance to oxidative stress, and that would result in improved implantation. (Methods and Materials) Primary
HESCs cultures were established, propagated, and confluent cultures remained were decidualized with 8-br-cAMP(0.5 mM) and MPA (10-6M), and with or without various concentration heparin or 4 days. Hydrogen peroxide(H2O2) treatment was used as a source of ROS. The amounts of PRL in culture media were examined. Western
blot was performed for protein analysis. The level of apoptosis was analyzed by cell detection ELISA Kit. (Results)
Treatment with 8-Br-cAMP plus MPA pronounced stimulated PRL secretion which is known as a decidualizing
marker. Addition of heparin at various concentrations to decidualized HESCs caused a dose-dependent incrase of
PRL production. Decidualized HESCs treated with heparin dose-dependently decreased the cell death rate inducedby oxidative stress. Although heparin treatment on its own did not elevate the expression of FOXO1 on
non-dedidualized HESCs, heparin dose-dependently augmented MPA/cAmp-induced FOXO1 and Mn-SOD
expression. Furthermore, heparin elicits phosphorylation PI3K-signal transduction pathway in primary
decidualized HESCs. (Conclusion) These results demonstrate that heparin-treated decidualized HESCs acquired
resistance to oxidative stress by induced expression of FOXO1 and Mn-SOD, suggesting that heparin may improvethe uterine environment for successful implantation.

P-038

The role of 5'AMP-activated protein kinase (AMPK) in human endometrial stromalcells○Yasushi Kawano, Sinya Karakida, Yufuko Utsunomiya, Hisashi Narahara
Oita University, Oita, Japan

Objective: 5'AMP-activated protein kinase (AMPK) is a fuel sensing enzyme that responds to decreases in cellularenergy state as reflected by an increase in the AMP/ATP ratio. Our objective was to clarify the role of AMPK in
cultured human endometrial stromal cells (ESC). Materials and Methods: Human ESC were cultured, and the effectof IL-1, 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) on the production of interleukin (IL)-8 andmonocyte chemoattractant protein (MCP)-1 was examined. The supernatants were collected, and IL-8 and MCP-1
were measured by ELISA. The activation of AMPK was detected by Western blot analysis using
anti-phosphorylated AMPK antibody. This study was approved by the institutional review board. Results: IL-1
increased the production of IL-8 and MCP-1 by ESC in a dose-dependent manner. On the other hand,
IL-1-stimulated IL-8 and MCP-1 production was significantly decreased following treatment with AICAR [IL-8:
IL-1β (1 nM); 73.0±15.9 ng/mL, IL-1β and AICAR (1 mM); 46.7±6.0 ng/mL (p<0.01); MCP-1: IL-1β (1 nM);
8,136.3±1,394.6 pg/mL, IL-1β and AICAR (1 mM); 4,685.1±432.8 pg/mL (p<0.01)]. The phosphorylations of
AMPK were determined by treatment with AICAR. Conclusions: The inhibitory effect on the production of
IL-1-stimulated IL-8 and MCP-1 via a mechanism involving AMPK were observed. The results suggest that
AMPK may play an important role of anti-inflammatory effect resulting to reduction of IL-8 and MCP-1
production. It is suggested that AMPK may contribute to the modulation of endometrial local factor in implantation
processes.

279

●Poster Presentations

P-039

Pinopode Formation upon Progesterone, hCG, and Clomiphene Citrate treatment
M Kabir-Salmani, ○M Shahali
National Institute of Genetic Engineering and Biotechnology, Dept. Molecular Genetics, Tehran. Iran

Background: Induction of ovulation is one of the primary steps in assisted reproduction while considered as one of
the main reasons for endometrial inadequacy in luteal phase. Hormonal treatment with progesterone or humanchorionic gonadotropin (hCG) appeared to improve pregnancy rates following hormone therapy for ovarian
hyperstimulation and became a common practice. However, little is known about endometrium status upontreatment with these hormones. Thus, this study was designed to investigate the effects of some routinely usedhormones, on the pinopode formation. Materials and Methods: A total of seventy five fertile and infertile females
ranging in age between 21 and 36 years participated in our studies. Study participants underwent ovarian
hyperstimulation with a long protocol method of a GnRH agonist or clomiphene citrate (CC) or received no
hormonal treatment considered as control group. Participants in GnRH agonist- or CC-stimulated groups wererandomized into designed experimental groups receiving progesterone, estradiol valerate or hCG which areroutinely used to improve endometrial status. Results: Statistical analysis indicated that the area-related numericaldensity of pinopodes in biopsies taken from patients who received progesterone treatment as a luteal phase support
following GnRH agonist-stimulated cycles were significantly higher than those who received hCG treatment andthose who received no treatment as well as those in natural control group. Our findings also indicated that pinopodedensity in patients who received CC plus estradiol valerate was significantly higher than those who received CCplus progesterone or CC alone. Pinopode density in patients who received CC plus progesterone or CC weresignificantly decreased compared to that of the natural control group. Conclusions: Our data demonstrated thatprogesterone following GnRH agonist-stimulated cycles increased pinopode density while has no significant effectwhen combined with CC. Instead, estradiol valerate increased pinopode density when induction of ovulation waswith CC. Moreover, it was shown that pinopode formation was reduced in stimulated cycles with CC while datafrom induction of ovulation using GnRH agonist protocol did not show this effect.

P-040

IS THERE ANY CORRELATION OF UTERUS POSITION AND DEEPTHEMBRYO TRANSFER ON PREGNANCY RATE IN IVF○Agus Supriyadi
Harapankita hospital, Republic of Indonesia

Introduction: To evaluate is there any correlation of uterus position and deepth embryo transfer on pregnancy ratein IVF. Material and methods: Cohort observational analytic study with retrospective data obtained from medical
record of patients participated in IVF programme at Harapan Kita Maternity Hospital Jakarta from January 2008 to
December 2008. Results: There were 50 patient, pregnancy 19 (38%) and no pregnancy 31 (62%). Position of
uterus: anteflexcy 29 patient (pregnancy 34.5%) and retroflexcy 21 patient (pregnancy 42.9%). Mean of deepth
embryo transfer on pregnancy success was 7.1 cm (SD 0.59), and unsuccess pregnancy is 6.98 cm (SD 0.63). Meanof the serum level estradiol was 1,450.98 ng/dL for pregnancy success, and 796.27 ng/dL for unsuccess pregnancy.
There were no significant correlation between uterus position and deepth embryo transfer with pregnancy rate
(p=0.547); (p=0.506). There was significant correlation between serum level estradiol and pregnancy rate (p=0.04).
Conclusion: there were no significant correlation between uterus position and deepth embryo transfer withpregnancy rate. Furthermore, there was significant correlation between serum estradiol levels with pregnancy rate.

280

●Poster Presentations

P-041

Influence of antisperm antibodies (ASA) on the outcome of infertility treatment○Yasufumi Shimizu, Takeshi Yorimitsu, Hiroshi Motoyama, Motohiro Ohara, Toshihiro Kawamura
Denentoshi Ladies Clinic, Kanagawa, Japan

Introduction: There has been suggested that antisperm antibodies (ASA) can impair the fertilizing capacity ofhuman spermatozoa, acting negatively on sperm motility and cervical mucus penetration, and at the level of in vitro
gamete interaction. The literature demonstrates that the various previously used treatments for immunologicalinfertility, i.e. medical therapy, intrauterine insemination with husband's spermatozoa (AIH) and in vitro
fertilization and embryo transfer (IVF-ET), usually had poor success. The purpose of this study was to investigate
the influence of ASA on the outcome of infertility treatment.Materials and Methods: We treated 28 couples whose
female partners were ASA positive and 13 couples whose male partners were ASA positive during 2007-2011.
Clinical results of ASA positive couples were compared.Results: First, we compared the results of IVF-ET (notincluding ICSI) between ASA positive females (5 cycles) and tubal factor (118cycles). Both female groups wererestricted under 35 years old in order to remove the influence of oocyte aging. Clinical pregnancy rate (gestationalsac+) per embryo transfer of ASA positive females was 100%, and that of tubal factor was 37.0 % (n.s.).
Fertilization rate of ASA positive female was 61.3%, and that of tubal factor was 74.7 % (n.s.). Abortion rate of
ASA positive female was 0%, and that of tubal factor was 25.9 % (n.s.). Next, we compared the results of IVE-ET(including ICSI) between ASA positive females (27 cycles) and ASA positive males (8cycles) in all ages. Clinicalpregnancy rate (gestational sac+) per embryo transfer of ASA positive females was 33.3%, and that of ASA
positive males was 33.3 % (n.s.). Fertilization rate of ASA positive females was 71.2%, and that of ASA positive
males was 68.8 % (n.s.). Finally, we compared the results of AIH between ASA positive females (39 cycles) and
ASA positive males(19cycles) and ASA negative couples (14602 cycles) in all ages. Clinical pregnancy rate
(gestational sac+) of ASA positive females was 2.6%, and that of ASA positive males was 5.3%, whereas that of
ASA negative couples was 7.3%, which was not significantly different. Conclusions: Our results suggested thatconventional IVF-ET was effective to ASA positive females, and pregnancy rate of IVF-ET was not different
between ASA positive females and ASA positive males. AIH was also effective to both ASA positive females and
ASA positive males.

P-042

The effect of ovarian stimulation on expression level of E-cadherin, Leukemia
inhibitory factor, Progesterone receptor and αvβ3 Integrin genes in mouse
blastocysts○Bahar Movaghar, Saeedeh Askarian
Royan Institute for Reproductive Biomedicine, ACECR, Tehran, IranIntroduction: The appropriate interaction between blastocyst and endometrium is essential for a successful
implantation. There are many factors involved in cross talk between embryo and endometrium including hormone
receptors, cytokines, adherence molecules and et ct. there are studies which show lack of these genes can be
resulted in implantation failures. In this study the expression level of some of these genes in superovulated mouse
blastocysts was compared with their expression in blastocysts from natural cycle. Materials and Methods: About
20 Female NMRI mice were injected with 7.5IU pregnant mare serum gonadotropin and the uteruses were flushed
3.5 days after given human Chorionic Gonadotropin and about 120 blastocysts were obtained from. Also 60 female
non stimulated mice mated with males and natural cycle blastocysts were flushed out from the uteruses 3.5 days
after mating. Expression level of E-cadherin, Leukemia inhibitory factor, Progesterone receptor and αvβ3 Integrin
genes was examined in 120 blastocysts obtained from hormone treated mice and in 120 natural cycle blastocysts by
Quantitative real time PCR method. Student t-test was used for data analyzing. Results: Results revealed that theexpression level of all studied genes were significantly lower in the hormone treated group than the natural cycle
blastocysts (p<0.05). However the highest decreasing rate was seen in β3 gene and the lowest change was seen in
αv gene. Conclusion: Although ovarian stimulation in ART cycles is utilized to obtain more oocytes, it may hassome disadvantages by down regulation of some genes involved in implantation process in blastocysts.

281

●Poster Presentations

P-043

Usefulness of elective single blastocyst transfer in the first treatment cycle forinfertile women aged less than 35.
○Hiroaki Shibahara, Shoko Hashimoto, Shiho Nagayama, Kazuhiko Shimada, Tatsuya Suzuki,
Mitsuaki Suzuki
Department of Obstetrics and Gynecology, School of Medicine, Jichi Medical University, Tochigi, Japan

Purpose: The modern trend of transferring less number of embryos is based on the idea that the satisfactorypregnancy rate (PR) could be maintained and the multiple pregnancies should be avoided. For such purposes,
elective single blastocyst transfer (eSBT) might be a useful treatment. However, it is still unclear that eSBT iseffective for both of younger women with relatively higher potential of conception and elder women with relatively
lower potential of conception. The aim of the present study is to know the indication of eSBT in the first treatment
cycle for infertile women. Method: Between Aug. 2006 and Dec. 2010, 124 infertile women were treated by the
first single embryo transfer following IVF or ICSI using fresh ejaculated sperm in our institute. Results: The
overall clinical PR was 33.0% (41/124). There were no multiple pregnancies. The PRs were 42.2% (27/64) inwomen aged less than 35 and 23.3% (14/60) in women aged 35 and more. There was a significant difference
between them (P=0.02). As for eSBT, the overall PR was 31.0% (17/55). The PRs were 46.2% (12/26) in theformer and 17.2% (5/29) in the latter. There was also a significant difference between them (P=0.02). The PR(33.3%) by day2/3 embryo transfer was better than that (16.7%) by day 4/5 embryo transfer in the elder women(P=0.13). Conclusions: These findings suggest that eSBT seems to be useful in the first treatment cycle for infertile
women aged less than 35. However, the PR by eSBT in the first treatment cycle for women aged more than 35 wasdisappointing. For such women aged 35 and more, day 2/3 single embryo transfer might be useful at least in thefirst treatment cycle. Alternatively, transferring of two embryos could be allowed for such women. Further study isrequired why some embryos derived from elder women are not suitable for longer culture in vitro.

P-044

A Summary of The clinical outcome of minimal stimulation IVF/ ICSIYan Ping Ma, Ze Wu, Bo Deng, ○Yonggang Li
Department of Reproduction and Genetics, Reproductive Medicine Centre, The First Peoples Hospital of Yunnan Province,
KunMing,China

In order to reach the goal of reducing the treatment cost, avoiding the risk of OHSS and multiple pregnancy,
gaining full-term single live birth. Natural cycle, minimal stimulation IVF and vertrification frozen, single
blastocyst transfer have been applied in Reproductive Medical Centre of the First Peoples Hospital of Yunnan
Province since 2009. This summary prospectively studied 28 natrual IVF cycles, 1065 minimal stimulation cycles,
435 fresh embryo transfer cycles and 182 frozen embryo transfer cycles from March 2009 to September 2010,
blastocyst formation rate was 46%, accumulative pregnancy rate was analyzed. The average age of patients innatural cycle was 30.8&plusmn 3.3 years. In 28 single blastocyst transfer cycles, the pregnancy test positive rate
was 52.6% (15/28), the clinical pregnancy rate was 42.9% (12/28). The average age of patients in minimal
stimulation cycle was 33.5 ± 3.8 years. Fresh embryo transfer was performed in 435 cycles. The pregnancy test
was positive in 184 cycles (42.3%, 184/435), the clinical pregnancy was achieved in 178 cycles (40.9%, 178/435).
In frozen embryo transfer cycles, pregnancy test positive rate and clinical pregnancy rate was 48.4% (88/182) and44.5% (81/182) respectively.

282

●Poster Presentations

P-045

Ovarian Response and Reserve Evaluation in consecutive IVF/ICSI-ET cycles○Mujdegul Karaca, A. Sertac Batioglu, Murat Ozel, Selcen Bahadir, Simla Karaca, Beril Gurlek
Dr ZTB Women's Health Hospital, Ankara, Turkey

Objective: To evaluate if IVF/ICSI-ET effects ovarian response and reserve in consecutive cycles and to put
forward it's clinical outcome. Methods: It has been retrospectively investigated 329 patients who had more than 3
IVF/ICSI-ET cycles between 2000-2008 among 5000 patients applied to our centre. Patients age, basal hormone
level, leading follicule count on day 6-7, gonadothropine dose applied, oocyte and embryo numbers gained andpregnancy outcome have been noted. Patients divided in to 4 age groups as less than 30 (129) group 1,between
30-34 (78) group 2, between 35-40 (129) group 3 and more than 40 (6) group 4. Results: Although basal hormone
level and number of oocytes and embryos gained isn't different, it has been found a significant rise ingonadothropine dose to get same result in each group. However leading follicule number decreased in consecutive
cycles for group 1 and 2, there wasn't a significant difference for group 3 and 4. After third cycle pregnancy andlive birth rates were increased. Conclusion: Although the rise in required gonadothropine dose to get the same
result in consecutive cycles is pointing a decrease in ovarian response; it doesn't show an ovarian reserve diminish.
But leading follicle number decline of group 1 and 2 in repeating cycles is indicating ovarian reserve downward for
ART patients especially younger than 35 years. It's thought-provoking that why ovarian reserve seems to be
reduced in younger patients and not in older ones for repeating cycles. This clinical outcome could be due tonaturally decreased ovarian capacity by age.

P-046

Our 11-year experience with sperm cryopreservation for patients with malignant
disease○Syuichi Iida, Masakuni Suzuki, Kazuhiro Hirayama, Ikuo Tachibana, Masahiro Katou,
Kouhei Tanaka
M. Suzuki's Memorial Hospital, Miyagi, Japan

Introduction; In recent years, damage to reproductive function has been a frequent and well-documented side effectassociated with the treatment of malignant disease. Because the gonadotrophic effect of a drug depends on its type,dose and the number of treatment cycles, it is impossible to predict which patients will have normal
spermatogenesis and which will remain azoospermic after treatment. Sperm banking before starting treatment for amalignant disease is currently considered the most effective method for preserving fertility. The aim of this study
was to describe the state of cryopreservation for each year of the study period, the profiles of patients withmalignant disease who wished to cryopreserve sperm, the conditions of the stored cryosperm, and the usage
conditions of the stored sperm in our hospital. We also analyzed various issues related to our 11 years ofexperience with banking sperm. Methods & patients; The number of sperm cryopreservations each year between
1999 and 2009 was determined. Then, the characteristics of forty-one patients with malignancy who desired tobank sperm were recorded; these characteristics included the patients'age, marital status, type of malignant disease,sperm condition at the time of cryopreservation and usage condition of the cryopreserved sperm. Results; The
sperm of 88 % (36/41) of patients was successfully collected and cryopreserved, yielding a total of 45 semen
samples. The number of new cryopreservations per year is currently stable. Most of the patients were young, single
and childless. Half of the patients were diagnosed with testicular tumors. Nearly half of the cryopreserved sperm
had counts >40 million/ml and a motility >50%. With respect to the usage condition of the cryopreserved sperm
samples, 36 % (16/45) of the samples are still being stored, 38 % (17/45) correspond to the samples of patients
missing in the follow-up and 27 % (12/45) had been used for ART or had been disposed of. The number ofaccumulated samples has gradually increased. Conclusion; Important factors for long-term sperm banking are thefollow-up with patients after cryopreservation and the continuity of sperm banking facilities. Thus, a close
connection between the sperm banking facility and the hospital at which the patients are being treated and followed
patients with malignant disease is important. Furthermore, it is preferable that sperm banking facilities should be
public large-scale clinics rather than private clinics.

283

●Poster Presentations

P-047

A Successful pregnancy and live birth after intracytoplasmic sperm injection withcryopreserved limited numbers of spermatozoa stored in the practical container○Yuji Endo, Yoshitaka Fujii, Hiroaki Motoyama
Kurashiki Medical Clinic, Okayama Japan

Introduction: Conventional freezing procedures are not suitable for a few spermatozoa. Some authors have attempted touse various types of containers such as zona pellucida for the cryopreservation of small numbers of spermatozoa.
Unfortunately, these containers are the only currently available options and they cannot be used universally. In ourprevious study (Endo et al, J Mamm Ova Res 2011), we have established a novel simple freezing technique for a single
spermatozoon using Cryotop (Kitazato Biopharma, Japan) and Cell Sleeper (Nipro, Japan), which are available
commercially and used universally. The aim of this study was to evaluate the efficiency of performing intracytoplasmic
sperm injection (ICSI) using sperm stored in these containers. Materials and methods: Spermatozoa were frozen in two
different containers, Cryotop or Cell Sleeper. Cryotop consists of a fine polypropylene strip attached to a plastic handle
and is equipped with a cover straw, and Cell Sleeper is a vial type of cell-freezing container, which equipped with an
inner tray and is sealed with a screw cap. Individual sperm were transferred to the containers using the ICSI pipettes
equipped with a micromanipulator. Immediately they were frozen in accordance with our method and stored in the
cryotanks. In the day of oocyte retrieval, the frozen sperm were thawed and retrieved. After following ICSI, a part of theoocytes were activated by 10 IU ml/ml of calcium ionophore A23187 (Sigma, USA) for 15 min. Fertilized embryos were
cultured until day 5, and single embryo transfers were performed in all cycles. Results: A total of 192 spermatozoa
obtained from six patients were frozen in 25 containers (7.7 cells per container). Two couples were underwent ICSI
program and stored sperm (19 cells) were thawed. Sixteen sperm were successfully retrieved and 3 of them were lost.
Sperm were injected individually into matured 12 oocytes and unused 4 retrieved-sperm cells were re-cryopreserved.
Fertilization was observed in 8 (67%), and all zygotes were cleaved. After embryo transfer, one woman was resulted insingleton pregnancy and concluded with full term delivery of healthy boy (2632g). Conclusions: Our clinical data showsa rare case of a successful delivery after transfer of a blastocyst derived from ICSI using the limited numbers of sperm
stored in the practical containers. Having a method of reliable sperm storage for severe male factor patients may reduce
multiple testicular surgical operations.

P-048

An evaluation of a modified slow cool cryopreservation technique and the efficacy
of a graduated estrogen regimen vs. natural cycles on pregnancy rates following
frozen embryo transfer○Jung. K Choe, Jerome H Check
UMDNJ, Robert Wood Johnson Med. School at Camden, Cooper Hosp./Univ. Med. Cntr., Dept OB/GYN, Div. Repro. Endo. &Infertility, Camden, New Jersey, U.S.

Introduction: Many in vitro fertilization-embryo transfer (IVF-ET) centers do not seem to have high pregnancy rates
following the transfer of embryos that have been cryopreserved using traditional slow cool methods. The present study
evaluated the efficacy of a simplified slow cool method that avoids what we believe is the weak point of the usual
method which in the planar freezer and instead uses an alcohol bath with slow cool controlled rate freezer (Biocool) witha one-step removal of the cryoprotectant. Furthermore the technique would compare the efficacy of natural vs. graduatedestradiol (E2) regimen on subsequent pregnancy rates (PRs).Materials and methods: The women were placed on the oral
E2 regimen. In general natural cycles were recommended if there was a contraindication for E2, e.g., history of breast
mass or cancer, thrombosis, migraine headaches or if the E2 regimen was started but was stopped because of side effects.
Embryos were either all cryopreserved at the 2 pronuclear phase in cases of risk of ovarian hyperstimulation syndrome orfor inadequate endometrial thickness. Otherwise the normal policy was to allow twice the number of embryos to cleaveto day 3 as intended to transfer and transfer the best half and freeze the rest. The embryos not allowed to cleave to day 3
were frozen at the 2 pronuclear stage. Results: Only 125 women were assigned to natural cycles vs. 1812 to thegraduated E2 regimen. The clinical PR per transfer was 33.6% with natural cycles vs. 42.5% with the E2 regimen(p=0.06, chi-square analysis). The live delivered PRs were 27.2% vs. 34.8% (p=0.10, chi-square analysis). The
respective implantation rates were 18.7% vs. 21.6% (p=0.25, chi-square analysis).Conclusion: The simplified slow coolcryopreservation protocol using a Biocool alcohol controlled rate freezer in lieu of the planar freezer with a one-step
removal of the cryoprotectant resulted in a very adequate pregnancy rate with frozen-thawed ET. The policy is to use up
all embryos and thus these data included transfers that were sometimes predominantly using embryos that werede-selected or even twice frozen twice thawed. There was a trend for higher clinical pregnancy rates using the graduatedE2 protocol. One must weigh the much increased use of the E2 hormone replacement not merely during the frozen ET
cycle but during the first trimester against the possible slightly lower pregnancy rates using a natural cycle.

284

●Poster Presentations

P-049

The effect of vitrification on Histone Modification of regulatory regions of H19, Igf2
and Mest imprinted genes in mouse embryo
1

1

2

3

○Bahar Movaghar , Saeideh Sahraei , Maryam Shahhoseini , Ali Farrokhi
1

2

Department of Embryology, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran, Department of Genetics, Royan
Institute for Reproductive Biomedicine, ACECR, Tehran, Iran, 3Department of Stem Cell, Royan Institute for Reproductive
Biomedicine, ACECR, Tehran, IranIntroduction: The effect of embryo vitrification and warming on epigenetic status of preimplantation embryo has tobe evaluated. Histone modification, as an epigenetic modification system, can control gene expression. Imprinted
genes H19, Igf2 and Mest are involved in regulating proper embryo development and imprinting disorders of thesegenes can be associated with some syndromes. This study was investigated gene expression level and modificationof three histone markers in transcriptional regulatory regions of these genes in vitrified-warmed mouse embryos.
Materials and Methods: Fresh blastocysts from NMRI mice were considered as control group. Also
Vitrified-warmed and non vitrified 8-cell embryos cultured into blastocyst stage were mentioned as experimentalgroups (I) and (II) respectively. Expression level of H19, Igf2 and Mest and also some histone modifications inregulatory regions of these genes were studied in the three groups. To gene expression evaluation, Real time PCR
was carried out on about 105 embryos in each group. On the other hand, acetylation of histone3 at lysine9 andmethylation of histone3 at lysine4 as gene activators and methylation of histone3 at lysine9 as a gene suppressor
were evaluated by ChIP assay method on 120 blastocysts in each group. REST test and ANOVA were used to data
analysis. Results: According to our study the expression level of all genes in group II was decreased significantly incomparison with control group (P<0.01). In group III only H19 and Igf2 had significant decreasing in comparison
with fresh blastocysts (P<0.01). Significant increasing in di-methylated histone3 at lysine9 was seen in theregulatory regions of all genes in vitrified group (P<0.01). Also tri-methylated histone3 at lysine4 and acetylated
histone3 at lysine9 were decrease in some regulatory regions but not all of them (P<0.01) in comparison with
control group. Additionally, di-methylated histone3 at lysine9 in regulatory regions of Igf2 and H19 but not Mest
was significantly increased in group III. Conclusion: These results indicate that vitrification and culture condition
can down regulate and modify histone pattern of regulatory sequences of H19 and Igf2 in blastocysts. Total histone
pattern in each group was coordinated with gene expression changes. Among different modifications,di-methylation of histone3 at lysine9 has the most coordination with gene expression changes in different groups.

P-050

Withdrawn285

●Poster Presentations

P-051

Which is more effective for pregnancy freezing embryos: in day-2, day-3 stage or
blastocyst?
○Tomio Sawada, Kaori Yoshikai, Sayumi Hori
The sawada women's clinic Nagoya reproduction center, Aichi, Japan

Introduction: In IVFand ICSI, the pregnancy result of frozen embryo transfer has improved compared with the
fresh embryo transfer in recent years. Moreover, it can be expected that the risk of ovarian hyperstimulation can be
reduced when all embryos are frozen. When all embryos freeze, the optimal developmental stage for cryopreseving
embryos remains controversial. Materials and Methods: All embryos freezing in our clinic are either frozen does atday-2,3 and then remainder is frozen by blastocyst. Thawed embryo transfer the first time replaces early day-2,3embryo or in some cases by blastocyst. When the first trial is not successful, the left embryo is transferred byblastocyst. The appropriate transfer protocols at the natural period or the hormone replacement is selected. It wasdetermined whether day-2,3 stage or blastcyst was more useful by this protocol. Results: Thawed embryo wastransferred in 41 cases and in 79 cycles and the pregnancy was a success in 24 cases and in 25 cycles (58.5% percases/ 31.6 % per cycle) in total. Ten out of 35 cases that transferred the first time day-2,3 early embryo reachedpregnancy (28.6%). One out of six cases that transferred the blastocyst the first time reached pregnancy (16.7%).
The second transfer attempt reached pregnancy in one out of 25 non-pregnancy cases in the early stage embryo. 24
cases were transferred the second time by blastocyst (11 cases early embryo to grew blastocyst among these) and11 cases (five cases in the growth blastocyst) reached pregnancy (45.8%). Two cases reached pregnancy in fivenon-pregnancy cases in the second transfer by the early embryo in blastocyst at the initial transfer (40%). The E2/P
ratio in the case that succeeded in a pregnancy by the second time compared with the first time pregnant cases,
there was no significant difference. Six cases ended in miscarriage within 25 cycles of the pregnancy. (24.0%)
Conclusions: Early stage day-2,3 and blastocyst are acceptable when all embryos freezing are executed at thefrozen time of the embryo. In the second transfer, it proved better that thawed early stage embryo were used in thetransfer after growing to a blastocyst.

P-052

Highest liquid nitrogen quality for vitrification process : Micro bacteriologicalfiltration of LN2
1

2

3

4

5

Ana Cobo , ○Damia Castello , B Weiss , C Vivier , Angel De La Macorra , F Kramp
1

2

3

6

4

IVI clinica de Fertitidad, Valencia, Spain, IVI clinica de Fertitidad, Air Liquide Head Office Industrial Merchant, Air Liquide Sante
International, 5Air Liquide Medicinal, 6Soprodi S.A.S., Quai du canal, Spain

Objective: To evaluate any risks of contamination of microorganisms in liquid nitrogen using Ceralin Filter
Equipment. Design: Analisys on-site (Octapharma plant) and in-situ (laboratory) of microorganism in LN2 forvalidation of the filter Ceralin equipment. Materials and Methods: Ceralin Filter Equipment is ceramic membranemade of multiple layers, formed into a multi-channel element.pore size of the ceramic membrane is 0.1 μm ; thus
the Ceralin can be considered as a sterilizing filter in accordance to FDA Guidelines on Aseptic Processing 1987:"A sterilizing filter produces a sterile effluent when challenged with the microorganisms Pseudomonas diminuta at
a given concentration (107 UFC/cm2 of filtering surface)" The on-site validation procedure consists of carrying out
sampling and analysis campaigns upstream/downstream from the filter. This analisys of samples was made using
impaction on agar-agar : bioimpactor MistralTM. with a flow 100 to 300L/min and a volume of sample of 200L.
Results: A significant reduction of microorganisms has been achieved by the Ceralin filter on-site. Guidelines forfilter validation (integrity tests) are not described nor performed in cryogenic conditions. Thus, correlations
between integrity test, bacterial challenge and particle challenge in standard conditions and in liquid nitrogen havebeen investigated. These studies lead to the strict specifications for Ceralin (only membranes whose bubble pointvalue is higher 2.2 bar). Validation of the filter efficiency in-situ has been done and can be done by customer whenwanted. Conclusions: Ceralin equipment offers a solution for a safer vitrification process. It guarantees nocontamination from liquid nitrogen and go onwards potential regulation imperatives related to this process.

286

●Poster Presentations

P-053

Comparing developmental potential and ultra structure of human blastocysts after
re-vitrification
1

2

1,2

○Mansoureh Movahedin , Zahra Alamolhoda , Nasim Ghorbanmehr , Mojgan Moradkhani

31

Tarbiat Modares University, Tehran, Iran, 2Shayamehr IVF Center, Tehran, Iran, 3Khatamolanbia Hospital, Tehran, IranIntroduction: Vitrification has shown to be a suitable and effective method of cryopreservation, but little is known
about re-vitrification of human embryos. The purpose of this study was to compare the developmental potential andultra structure of human blastocysts after re-vitrification and determining the best stage of human forre-vitrification. Materials and Methods: 95 cleavage embryos were obtained from the patients who didn't want tohave embryo freezing because of different reasons. After getting informed constant, the embryos were divided into
two groups. In the first group, 42 embryos (4-8 cell stage embryos) were vitrified and warmed using the Cryotop
technique. After evaluation the survival rate, the alive embryos were vitrified and warmed again with the same
method. After thawing, the survival rate was assessed and the alive embryos were cultured for 3-4 days and in theend of cultivation, the blastocyst rate was determined. In the second group, 53 embryos (4-8 cell stage embryos)
were vitrified and warmed with the above mentioned method. After warming and evaluation their survival rate, thealive embryos were cultured for 3-4 days and then the blastocyst rate was evaluated. The obtained blastocysts werere-vitrified and warmed and the survival rate was assessed. Ultra structure of the obtained blastocysts in both group
after re-vitrification were studied, too. Results: There was not significant difference between survival rates of twogroups after vitrification-warming (92% vs. 89% respectively for the first and second groups). 20 out of 47
embryos (43%) of the second group reached to blastocyst stage after thawing. The survival rate in the first group
after re-vitrificatin was 89.7%, meanwhile it was 30% in the second group and there was significant difference(P<0.05). The blastocyst rate after re-vitrificatin in the first group was 25.7%. Ultra structural study revealed
disorganization of cytoskeleton of the blastocysts after re-vitrification in the both groups. The regularity of
mitochondria was also disappeared and some vacuoles were seen in the cytoplasm of re-vitrified blastocysts.
Conclusion: The results showed that re-vitrification of cleavage embryos has less harmful effects comparing to
re-vitrification of blastocysts. The reason for delay of blastocyst formation could be ultrastructural changes
following re-vitrification.

P-054

Examination of blastocyst transfer after pronucleate embryos were slowly
freeze-thawed○Hiroei Ohhashi, Yukio Ohmomo, Osamu Arakawa
Arakawa Ohmomo Angel Mother Clinic, Niigata, Japan

Objective: We examined the usefulness of blastocyst transfer after pronucleate embryos (2PN) were slowly
freeze-thawed. Subjects: One hundred and four cycles in 98 patients who received slowly freeze-thawed 2PN wereinvolved in this study from January through December 2010. Freezing was applied to the following cases: patientswith multiple conception failures, Clomifene citrate in poor-responders, for the prevention of OHSS, when ET falls ona holiday, there are surplus embryos available, etc. Methods: When freezing, a blastocyst was exposed to 20% serum
PBI for five minutes, to 1.5 M PROH for 10 minutes, and to 1.5 M PROH+0.1 M sucrose for 10 minutes, then placedin an alcohol bath at 10degree celsius in a programmable freezer. When thawing, straws were left to stand at room
temperature for 40 seconds after being taken out of liquid nitrogen, then dipped in water at 23degree celsius for 10
seconds, and transferred into a thawing solution. The thawing solution comprised 1.0 M PROH+0.1 M sucrose
decreasing by 0.25 M / 5 minutes and the specimen was cultured for five days after being dipped in 0.1 M sucrose and
PBI for 5 minutes, respectively. The embryos transplanted it in a HRC. Results: The 2PN survival rate was 96.7%(540/558), the cleavaged rate was 99.0% (535/540), the rate of blastocyst formation for D5 was 31.2% (169/540), andblastocyst transfer became possible for 74% (77/104) of frozen embryos in the thawing cycle. The pregnancy rate
after blastocyst transfer was 55% (39/77), the pregnancy rate in the thawing cycle was 39% (41/104), and the abortion
rate in the thawing cycle was 26% (11/41). The pregnancy rates after blastocyst transfer and in the thawing cycle
classified by age were 83% (5/6) and 83% (5/6) in the 20s, 60% (15/25) and 46% (15/32) between 30 and 34 years
old, 56% (14/56) and 48% (16/33) between 35 and 39 years old, 50% (5/10) and 29% (5/17) between 40 and 42 years
old, and 9%(1/11) and 6%(1/16) for over 43 years or older, respectively. The pregnancy rates after blastocyst transfer
and in the thawing cycle classified by the applied conditions were 66% (4/6) and 36% (4/11) for multiple conception
failures, 16% (2/12) and 15% (3/19) for Clomifene citrate, 66% (20/30) and 58% (20/34) for the prevention of OHSS,
62% (5/8) and 55% (5/9) for ET days falling on a holiday, 62% (5/8) and 60% (6/10) for freezing of surplus embryos,and 30% (4/13) and 22%(4/21) for others.Conclusion: In this examination, it is thought to be effective in a case 40
years or younger.

287

●Poster Presentations

P-055

5/-(N-Ethylcarboxamido) adenosine (NECA) improves angiogenesis intransplanted human ovarian tissue: potential for fertility preservation
1

2

3,4

5

○Maryam Hormozi , Saeed Talebi , Amir Hassan Zarnani , Mahmood Jeddi-Tehrani ,62Ladan Hosseinigohari , Mohammad Mehdi Akhondi
1Biochemistry Department, lorestan university of medical sciences, Khorramabad,Iran, 2Reproductive Biotechnology Research Center,
Avicenna Research Institute, (ACECR), Tehran, Iran, 3Nanobiotechnology Research Center, Avicenna Research Institute, (ACECR),
Tehran, Iran, 4Immunology Research Center, Tehran University of Medical Sciences, Tehran, Iran, 5Monoclonal Antibody ResearchCenter, Avicenna Research Institute, (ACECR), Tehran, Iran, 6Biochemistry Department, Tehran university of Medical Sciences,
Tehran, IranIntroduction: In the past 3 decades by enormous progress in cancer therapy the possibility of successful recovery
from this disease has enhanced. However, many of cancer patients become infertile after treatment and this can be
an important concern for people at reproductive age. There are limited ways to restore fertility of womenpost-treatment such as cryopreservation of embryo, oocyte and ovarian cortex. One of the promising methods torestore fertility ability of these patients is ovarian tissue banking (OTB). OTB is based on the isolation of fragmentsof cortex of ovary before cancer therapy, freezing and transplanting them after cancer treatment.There are many
drawbacks such as apoptosis and considerable reduction of follicular density in the transplanted ovary. Many
studies have shown that these problems mainly result from ischemic damages and not from freezing/thawing
process of the tissue. One solution to reduce ischemic damage is enhancing angiogenesis after transplantation ofovarian cortex tissue. Based on the angiogenesis capacity of adenosine, the aim of this study was to evaluate theangiogenesis potency of NECA, and adenosine agonist, in triggering angiogenesis of transplanted human ovariantissue. Material and Methods: Human ovarian tissue fragments were transplanted to nude mice. Four hours posttransplantation (PT), mice were treated with NECA, while control group received only vehicle. The transplanted
fragments from each group were recovered on days 2, 7 and 30 and studied for expression of Angiopoietin1,
Angiopoietin2, VEGF-121 and VEGF-189 at both gene and protein levels (by real time PCR and western blot
respectively) and vascular density. RESULT(S): On the 2nd day PT, the level of Ang1 gene expression was
significantly lower than that in controls, while the opposite result was obtained for VEGF-189. These results werealso confirmed at the protein level. The differences were, however, not statistically significant for Ang2 and
VEGF-121. The density of vessels in NECA group elevated significantly at day 7 PT compared to pre-treatment
state.CONCLUSION(S): Since the process of angiogenesis is more critical in the first 48 hours PT, our results
suggest a beneficial role for NECA in this process.

P-056

Assessing risks of birth defects in ART babies○Coralia V. Stefanescu, Ion M Rusa, George Costea, Lenuta Malcea
Euromaterna Hospital, Constanta, Romania

The incidence of congenital malformations in infants born after reproductive technology has been much discussed
in the literature with controversial conclusions. Slightly more than 4% of babies born via assisted reproductive
technology (ART) may have major birth defects. One of the explanation may be due to the higher rate of multiples
pregnancies which may stands for a higher rate of abnormalities. In this paper we try to overview the literature
figures and, also, we present a case of twin pregnancy, obtained by IVF, in which one of the fetuses was affectedby duodenal atresia. The pathology is due to failure of recanalization of this intestinal segment. Its frequency varies
between 1/2500 and1/10000 live birth and it has high risk of an associated chromosomal anomaly. The diagnosis
was based upon follow up ultrasound scans (recognition of the classic double bubble sign, associated withpolyhydramnios) and the prognosis was good after the surgical cure, despite the premature birth. As a fact, weconclude that councelling about possible risks associated with fertility treatment (including multiple pregnancy andfetal malformations) is an important part of the fertility treatment process and, therefore, obstetricians andpediatricians, as well as certified infertility counselors, need to become sources of an accurate information.

288

●Poster Presentations

P-057

Two cases of ovarian abscess after oocyte retrieval○Aki Oride, Haruhiko Kanasaki, Kohji Miyazaki
Shimane University School of Medicine, Shimane, Japan

Ultrasound-guided transvaginal oocyte retrieval has been known as he standard ovum pick up (OPU) method at invitro fertilization and embryo-transfer (IVF-ET). Pelvic abscess formation is one of the complications induced by
OPU. We report 2 cases of ovarian abscess which occurred after OPU. Case 1 was a 29 years old nurse. We
punctured and aspirated fluid from an endometrioma followed by OPU. Eleventh day after OPU, she developed
fever and bilateral ovarian swelling with an abscess-like mass observed by transvaginal ultrasonography. She washospitalized and underwent intravenous antibiotic treatment. Because of the continuing high inflammatory
reaction, we punctured and drainaged the ovarian abscess. MRSA was detected in the content fluid. By changing
antibiotic which targeted MRSA, her inflammatory reaction improved immediately. Case 2 was a 43 years oldhousewife. She developed fever and abdominal pain at second day after OPU. Her symptoms and the laboratory
findings improved immediate after the start of antibiotic treatment. MRSA should be considered as an etiologic
agent in cases of pelvic inflammation after OPU.

P-058

An experience of an infertility treatment for a woman with benign metastasizing
leiomyoma○Tsuyoshi Hashiba, Toshio Hamatani, Naoaki Kuji, Kou Sueoka, Yasunori Yoshimura
Keio University School of Medicine, Tokyo, Japan

Benign metastasizing leiomyoma (BML) is a rare disease in which solitary leiomyoma-like nodules are present in a
distant location. As BML is found primarily in women of reproductive age, the origin of BML is presumed to be
the uterus. Approximately 150 case reports of BML have been published. We report an experience of an infertility
treatment in a woman with BML. A 40-year-old nulliparous woman was referred to our University hospital for
surgical operation of the abdominal wall tumor and the uterine myoma. Past history includes an abdominal
myomectomy for the uterine myomas, which were diagnosed as cellular leiomyoma in the pathologicalexamination, 4 years prior to referral. On the first visit, the patient has a growing abdominal tumor of 6 cm
diameter and a uterine tumor of 6 cm diameter. After through workup, she underwent an abdominal tumor
resection and an abdominal myomectomy in our hospital. The pathological examination was reported as cellularleiomyoma of the uterus, metastasizing to the abdominal wall. As she may have the risk of tumor spreading or
metastasizing, furthermore having the right tubal occlusion, we decided to perform in vitro fertilization and embryotransfer for an infertility therapy. Under the long luteal protocol, 19 oocytes were retrieved, followed by moderate
ovarian hyperstimulation syndrome. The patient achieved a single pregnancy by a thawed embryo transfer innatural cycle, resulting in delivering a healthy infant by cesarean section, in which any developing tumors were notobserved. Before starting an infertility therapy in a woman with BML, making a diagnosis of BML precisely isessential. Meticulous sampling of the pathology specimen should be undertaken to exclude leiomyosarcoma, which
unlike BML, has an aggressive course.

289

●Poster Presentations

P-059

A Transgenic Mouse Model of Breast Cancer Using tissue specific MMTV
Promoter
1,2

1

1

2

1

○Maryam Shahali , Ehsan Ranaei , Shams-Ara Mahdi , Javad Mowla , Maryam Kabir-Salmani ,3
Karim Nayernia
1National Institute of genetic engineering and Biotechnology, Tehran, Iran & Tarbiat Modares University, Tehran, Iran, 2Tarbiat
Modares University, Tehran, Iran, 3GENOCELL CO.

Background: Piwil2, a member of AGO/PIWI family of proteins, has been reported to be expressed in
precancerous stem cells (pCSCs), tumor cell lines and various types of human cancers. In human breast cancer,
previous findings have been determined Piwil2 and its effector signaling pathways as key factors in the
proliferation and survival of breast cancer stem cells. To study the roles of Piwil2 in breast cancer, a transgenic
mouse model has been developed containing mouse mammary tumor virus (MMTV)-piwil2cDNA construct.
Methods: Suitable vector containing pMMTV-piwil2CDNA was constructed and linearzed by two desirable
restriction enzymes. Then, the MMTV-piwil2cDNA construct was injected into the pronuclei of 0.5-day NMRI
embryos. Results: Five transgenic founder male mice were produced as determined by southern blot and PCR
analyses. Four founder animals were able to transmit the transgene to their offspring. Conclusion: Tumorigenesis
effects of Piwil2 on the breast tissue in the F1 generation are being investigated.

P-060

Effect of genistein administration on intratesticular testosterone level andspermatogenesis in rats treated with busulfan○Heejun Chi, Kangwoo Cheon, Jonghyun kim, Giyoung Kim, Jaeseok Lee, Sungil Roh
i-Dream Center, Mizmedi Hospital, Seoul, Korea

Objective: To investigate the effect of genistein (isoflavon) administration on spermatogenesis recovery of the rats
treated with busulfan (chemotherapy drug).Materials and Methods: 185 male rats were assigned to Control,
Busulfan, Busulfan/GnRH agonist (-a), Busulfan/Genistein 50 mg, and Busulfan/Genistein 100 mg treatmentgroups. A single intraperitoneal injection of busulfan (25 mg/kg) was done to induce a chemical damage to testis.
Three weeks after the busulfan injection, an additional injection of GnRH-a (0.625 mg) or oral administration ofgenistein (50 mg or 100 mg) for 4 weeks was done. Thirteen weeks after the busulfan injection, serum andintratesticular testosterone (ITT) levels, weights of testes, and histological characteristics of the testes were
investigated. Results: Testosterone level in serum (2.70 ng/ml) was significantly reduced after busulfan injection(1.65 ng). Additional treatments of GnRH-a (1.11 ng) and genistein (50 mg: 1.52 ng, 100 mg: 1.71 ng) did notreverse the reduced testosterone level by busulfan. However, ITT level (Control, 67.7 ng / g-testis) was
significantly increased after busulfan injection (365.1 ng), and the increased ITT level was reversed by additional
GnRH-a (159.6 ng) and genistein 50mg (170.0 ng), 100mg (155.0 ng) treatments. Weight of testis (Control, 3.49 g)
was significantly decreased by busulfan injection (1.29 g), but the reduced weight of testis was partially recoveredby GnRH-a (1.77 g) or genistein (50 mg: 1.85 g, 100 mg: 1.70 g) treatment. Histological examination revealed thatbusulfan induced a severe damage to spermatogenesis, but the damaged spermatogenesis was partially recoveredby both additional GnRH-a and genistein treatments. Conclusions: Genistein enhanced the recovery of thechemically impaired spermatogenesis through the suppression of ITT level. Therefore, we suggest that genistein isa safe substitute for hormones to recover the impaired spermatogenesis of human cancer patients treated withchemotherapeutic drug, with less risk of side effects.

290

●Poster Presentations

P-061

The role of Doppler in predicting endometrial thickness, pattern and blood flow ininfertile women undergoing IVF cycle in a tertiary care institute○Anupama Bahadur, Neeta Singh, Neena Malhotra, Ashok Bhatt, Suneeta Mittal
All India Institute of Medical Sciences, New Delhi, India

Introduction: Endometrial receptivity is crucial to implantation of an embryo. 2D power Doppler was used toevaluate the association between endometrial thickness, its pattern and blood flow and pregnancy outcome inwomen undergoing in-vitro fertilization. Material & Methods: A total of 140 infertile women attending theinfertility clinic at All India Institute of Medical Sciences, New Delhi between January 2009 to December 2010
was included in this prospective study. The study was approved by the Institutes Ethics Committee. The cause ofinfertility in these patients was tubal factor, male factor, and unexplained infertility. All patients were stimulatedusing the standard long protocol. The endometrial thickness, pattern and blood flow was recorded on the day of
HCG administration. Results: The mean age was 34.5 years and mean duration of infertility was 7.6 years. 69% of
patients had primary infertility and 31% had secondary infertility. The mean endometrial thickness was 8.4mm
with endometrial blood flow in zone 1 in 38 patients, zone 2 in 44 and zone 3 in 58. 28.4% women with anendometrial thickness between 6.4 to 12mm achieved a successful pregnancy. Also a higher pregnancy rate wasobserved when the blood flow was in zone 3(49.2%) compared to zone 1 (16.7%). Conclusions: The endometrial
thickness and vascularity have a predictive value in implantation of an embryo in women undergoing assisted
reproductive techniques. However, the study sample was small to make a definite statement.

P-062

Reduced blastcyst formation rate in women in advanced reproductive age withhigh Anti-Mullerian hormone level○Takahiro Noda, Shinako Hashimoto, Mitsuru Iwamoto, Syuichi Iida, Kohei Tanaka,
Masakuni Suzuki
Suzuki Memorial Hospital, Miyagi, JapanBACKGROUND: Anti-Mullerian hormone (AMH) is widely thought to be a useful marker of ovarian reserve. It isalso known that serum AMH level declines with aging. However, there are some exceptional cases with still higher
serum AMH levels in advanced reproductive age. We focused on this group and studied the results of blastcyst
culture for IVF or ICSI. METHOD: We studied 35 cycles of IVF-ET or ICSI undergone at Suzuki memorial
hospital from September 2009 to February 2011. Prior to ovarian stimulation, serum AMH concentrations were
measured. These cycles are divided in three groups. Group A: high AMH level (>20 pmol/ml) - advanced age(>=36) group (n=6), Group B: low to normal AMH level (<20 pmol/ml) - advanced age (>=36) group (n=14),
Group C: younger age (<36) Group (n=15). Number of oocyte yield, fertilization rate, blastcyst forming rate werecompared. RESULT: Mean oocyte yield in group A was 10.5, which is significantly higher than that in group B(4.6) and C (5.9). No significant differences in fertilization rates were observed among three groups; Group A(56%), Group B (51%), Group C (54%). Blastcyst forming rates were as follows; Group A (14%), Group B (39%),
GroupC (54%). Significant reduced blastcyst forming rate in group A comparing with group C were noted(p=0.026). CONCLUSION: Although relative large number of oocyte yield may be expected, women inadvanced age with higher serum AMH levels were characterized by significant reduced blast cyst forming rate.

291

●Poster Presentations

P-063

Effects of salpingectomy on ovarian response in controlled ovarian
hyperstimulation for in vitro fertilization: a reappraisal
1○Israel Wagman , Benny Almog

1,2

1

1

1

, Ishai Levin , Gali Barkn , Dina Kovalsky , Togas Tulandi

2

1

Tel Aviv Sourasky Medical Center, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel., 2Department of Obstetrics andGynecology, McGill University, Montreal, Quebec, Canada

Objective: To evaluate the effects of salpingectomy on ovarian response in controlled ovarian hyperstimulation(COH). Setting: University-based tertiary medical center. Study design: Retrospective study Patients: 36 women
who underwent controlled ovarian stimulation cycles for in-vitro fertilization (IVF) before and after salpingectomy
Interventions: Laparoscopic salpingectomyMain outcome measures: The number of dominant follicles andoocytes aspirated before and after salpingectomy. We also evaluated maximal estradiol levels, duration of
stimulation and average daily dose of gonadotrophins. Results: The overall number of dominant follicles and thenumber of oocytes aspirated before and after salpingectomy were comparable (7.2 ±3.8 vs. 7.3 ±3.7 and 10.2 ± 6.6
vs. 10.3 ±7.4 respectively). Maximal E2 levels and the daily doses of gonadotrophins before (1899 ± 185 pg/ml
and 217.8 ±10.0 IU) and after surgery (1997 ±231 pg/ml and 239 ±16.3IU) were also similar. There was no
significant different in the number of dominant follicles before and after surgery on the operated side (3.8 ± 2.2 vs.
3.7 ±2.0). Regression analysis to assess the effect of unilateral versus bilateral salpingectomy showed no effect on
the main outcome measured. Conclusions: Unilateral or bilateral salpingectomy does not influence ovarian
response in controlled ovarian hyperstimulation.

P-064

Semen analysis and evaluation of ICSI outcome in patients infected with hepatitis
B virus
1

2

2

1

2

○Mounir Ajina , Neila Hannachi , Khaled Youssef , Souhir Mehri , Jalel Boukadida ,3
Ali Saad1

Unit of reproductive Medicine, University Hospitol Fahat Hached, Sousse, Tunisia, 2Laboratory of Bacteriology, University Hospital
Farhat.Hached, Sousse, Tunisia, 3Laboratories of Cytogenetic, Molecular Biology and Human Biology of Reproduction, Farhat HachedHospital, Sousse, Tunisia.

Introduction: Hepatitis B infection represents the most important virus to manage in assisted reproduction
techniques (ART) Tunisian candidates, screening of this sexually transmitted virus is recommended. The successof ART in couple infected with hepatitis B virus (HBV) was not clearly established. Several inflammatory andbiological disturbances exist in hepatitis B and interference with the biological steps of ART was possible. The aimof our study was to verify a possible correlation between infection with hepatitis B and sperm abnormalities andthe biological assessment of IVF. Materiel and Methods: This is a case-control study of 56 couples infected with
hepatitis B virus (cases) followed by consultations of the Unit of Reproductive Medicine of Sousse and candidates
for in vitro fertilization (IVF). HBV markers were tested for both partners in case of at least one positive forhepatitis B virus surface antigen (HbsAg). Their recruitment was done between February 2004 and February 2010.
These markers included hepatitis B surface antibody (anti-HBs) and Hbe antigen and antibody (HbeAg andanti-Hbe). Serologies were assessed by commercially available assays.Results: The HbsAg rate was significantly
higher in men 61% than in women 39%, p= 0.04. No serology HBeAg type was positive. Among the 59 couples
infected with hepatitis B s only 3.4% of couples were jointly infected. The most frequent sperm abnormalities inpatients with HBsAg was the asthenozoospermia, 62.5% vs 20% in the control group, P=0.001. The oligospermia
was significantly higher in the control group, p=0.005. Tubal obstruction was the most common abnormality
among infected women 95% vs 50% in the control group, P=0.008. The sperm bacteriology was significantly
higher positive in cases with a prevalence of 28.8% against 9.2% in control group, p= 0.01. The pregnancy rate
among cases and controls was equal respectively 12.1% and 16.7% with an insignificant difference. Conclusion:
The asthenozoospermia was the most common abnormality in infected man with hepatitis B virus. But in infected
women, the tubal obstruction was the most frequent anomaly and was associated with Chlamydia trachomatis.

292

●Poster Presentations

P-065

Effect of Chemotherapy on Spermatogenesis in Testicular Cancer Patients
1

1

1

2

1

○Miki Fuse , Takashi Imamoto , Takanobu Utsumi , Takumi Endo , Naoki Nihei ,1
Tomohiko Ichikawa
1

Department of Urology, Graduate School of Medicine, Chiba University, Chiba, Japan, 2Department of Urology, Toho University
Sakura Medical Center, Chiba, Japan[Purpose] We evaluated fertility status after treatment for Testicular cancer (TC) by conducting a large,
retrospective study. [Materials and methods] A retrospective study was conducted of 103 patients who wasdiagnosed unilateral gonadal germ cell tumor between 1983 and 2010 in our facility, and free of relapse. Semen
analysis and serum hormonal level (lutenizing hormone: LH, follicle-stimulating hormone: FSH, testosterone:
TST) were obtained for each patient, as well as the fertility follow-up. Seventy-six patents were treated withchemotherapy after orchidectomy. [Results] Thirty-seven patients were performed routine semen analysis before
chemotherapy. The mean sperm motility was below (40.1±28.3%), while the mean sperm concentration was above
the normal (27.0±33.0*106/mL). In non-azoospermic patients (n=32), fifteen were asthenozoospermic. After
chemotherapy, seminal parameters were recovered to the normal value within 30±31 months in twenty-two
patients. Patients treated with 5 and more cycles of chemotherapy were hard to recover the normal sperm condition.
With regard to the hormone level, FSH was kept high in the patients who didn,t recover the semen data, while TST
has been kept within the normal range. Of 21 patients who had attempted to achieve pregnancy with their partners,
nine patients had succeeded and 7 patients had children (60±39 months after treatment). In the patients who
succeeded to have children, sperm concentration and motility just before pregnancy were 53±36*106/ml and61±26%, respectively. [Conclusions] About half of the TC patients showed poor sperm quality before
orchidectomy. Our data demonstrated that the patients who recovered spermatogenesis after chemotherapy showed
improved sperm concentration above 20*106/ml within 2 years and recovered fertility. Patients treated with highdose chemotherapy or 5 and more cycles of PVB (cisplatin, vinblastine, and bleomycin) or BEP (bleomycin,
etoposide, and cisplatin) showed poor recovery of spermatogenesis, but their TST values were kept within normal
level.

P-066

Reproductive outcomes following laparoscopic myomectomy in patients withinfertility○Kohei Tanaka, Masakuni Suzuki
M. Suzuki Memorial Hospital, Miyagi, Japan

Objectives: To evaluate the outcomes of gasless laparoscopic myomectomy in infertile patients. Methods: We
conducted a retrospective clinical analysis of 170 patients who underwent laparoscopic myomectomy for infertility.
Gasless laparoscopic myomectomy was performed by lifting two K-wires inserted into the lower abdomen
subcutaneously. The patients were followed up for more than one year. We evaluated clinical findings, operative
findings and obstetrical outcomes. Results: One hundred ten of the 170 patients (64.7%) conceived after
myomectomy. Sixty-six (60%) received no treatment for infertility after the operation. Sixty patients (36.3%)
remained infertile, despite most (68%, 41/60) receiving ART (assisted reproductive technology). Average age andbody weight were significantly lower in the pregnant than in the nonpregnant group (33.7 vs. 36.7 years; 53.9 vs56.5 kg.). The mean duration of infertility before laparoscopic myomectomy was shorter in the pregnant than in the
nonpregnant group (3.4 vs 5.9 years). The size, location, number and weight of myomas did not differ between thetwo groups. The drop on hemoglobin concentration, operative time and concomitant surgeries did not influence
fertility outcomes. The rate of umexplained infertility factors was higher in the pregnant than in the nonpregnant
group (58/110, 19/60; p=0.008). Conclusions: Gasless laparoscopic myomectomy is useful, effective and safe forinfertile patients. This operative procedure is an important therapy for infertile patients with myomas regardless of
their location, number or size.

293

●Poster Presentations

P-067

The role of CINC/gro in the rat ovulatory process○Yu Tanaka, Akira Kuwahara, Yuya Yano, Yuri Yamamoto, Kenji Hinokio, Minoru Irahara
Deparatment of Obstetrics and Gynecology, The University of Tokushima, Institute for Health Biosciences, Tokushima, Japan

CINC/gro is a CXC-family chemokine similar to interleukin-8 in rats. CINC/gro is one of the factors that regulate
the ovulation process. However, the mechanism that regulates atresia of post-ovulation in ovaries is not clearly
defined. Therefore, we aimed to determine whether antibody-blocking of CINC/gro might change the number ofovulated oocytes and modulate neutrophil infiltration. We investigated the effect of antibody on the level ofinflammatory cytokines production, and on atresia of the follicles. Apoptosis was measured the TUNEL method
and by analysis of the mRNA expressions of Bcl-2 and Bax. Anti-CINC/gro antibody treatment decreased thenumber of ovulated oocytes. The mRNA levels of COX-2 and IL-1 beta were decreased by antibody treatment,
whereas that of TNF alpha was increased. TUNEL analysis indicated a large number of apoptotic cells in the
antibody group compared to control as well as a significant increase in the Bax/Bcl-2 ratio 24 h after hCGadministration. These findings suggest that ovulation is accelerated by neutrophil infiltration into the theca layer.
CINC/gro appears to synergize with IL-1 beta for ovulation. In contrast the data suggest that expression of
CINC/gro suppresses TNF alpha expression and that expression of CINC/gro therefore prevents follicles fromundergoing atresia and apoptosis.

P-068

A Commercially Available Dual-Buffered IVF Handling Medium Containing HEPES
and MOPS Maintains Stable pH and Supports Human Sperm Survival, Normal
Fertilization Following ICSI and Embryo Development1○Jason E Swain , Marlane
1

2

3

4

Angel , Nadir Cira , Juergen Liebermann , Thomas Pool2

5

3

University of Michigan, Ann Arbor, MI, Laurel Fertility Care, San Francisco, CA, USA, Bacchi IVF Center, Istanbul, Turkey,
Fertility Center of Illinois, Chicago, IL, USA, 5Fertility Center of San Antonio, San Antonio, TX, USA

4

Introduction: Maintenance of stable pH is important to reduce stress in the culture environment and optimize
embryo development. Buffers, like HEPES and MOPS, are used to stabilized pH of media used outside the
incubator. Current commercially available media utilize a single buffer. However, use of a single buffer limits theability to adjust optimal buffering capacity. For example, the pKa, or optimal buffering of MOPS at 20oC is 7.20,
while HEPES has a pKa of 7.55. At 37oC HEPES has a pKa of 7.31, while MOPS is 6.95. Furthermore, potential
toxicity concerns exist with elevated concentrations of buffers used in mono-buffered media. Therefore, media
utilizing a single buffer may not be ideal. Use of a dual buffered media containing MOPS and HEPES may permit
benefits of both buffers and provide a more appropriate buffering over a range of temperatures, while permitting
use of lower buffer concentration.
Materials and Methods: A dual-buffered medium containing HEPES and MOPS (pH SecureTM,Irvine Scientific),
was tested against control HEPES buffered media. pH of media was measured and compared. Standard QC
assays including a 24h HSSA and a 96h 1-cell MEA were used to assess media efficacy. Finally, human oocytes
were split between each media and held for ICSI and resulting fertilization compared. Data were analyzed using
Fishers exact test.
Results: The pH of pH SecureTM and control HEPES media were 7.27 and 7.33, and change in pH following 1
month were not significantly different (0.08 vs. 0.07), respectively. pH SecureTM maintained sperm motility at rates
equivalent to control media after 24h, resulting in a 14.8% and a 20.2% decrease in motility, respectively (n=5).
Following 2h exposure to pH SecureTM or control media, no differences in blastocyst formation (80% vs. 76.5%) or
hatching (45% vs. 28) were observed following 96h of culture (n=2). Finally, no differences in rates of normal
fertilization following ICSI were apparent in either pH SecureTM (78.6%) or control HEPES media (74.2%) (n=47patients, 594 oocytes).
Conclusion: Use of a dual-buffered medium containing HEPES and MOPS maintains a stable pH environment,
supports human sperm survival, normal fertilization following ICSI and mouse embryo development at rates
comparable to traditional HEPES buffered medium. Combining two buffers to regulate media pH offers thepotential advantages of reducing individual buffer concentration, as well as the ability to optimize buffering
capacity.

294

●Poster Presentations

P-069

Development of Mouse and Human Embryos in a Low Humidity Incubator○Chiemi Mori, Tadashi Okimura, Fumihito Aono, Yuji Takehara, Osamu Kato
Kato Ladies Clinic, Tokyo, Japan

Objective: Recently, a dry culture system has been designed - without a water tray - which could diminish the riskof any contamination. At ASRM2010 we have already reported that water evaporation and osmolarity changes
could be efficiently controlled in a dry culture system. The aim of the present study was to compare the efficacy ofdry and humid culture systems by evaluating blastocyst formation rates of mouse and human embryos. Materials
and Methods: Two separate incubators were used to compare embryo development. For the dry and humid
incubator system an EZ-Culture System and a conventional AP30 incubator were used respectively (ASTEC,Japan). 2,225 2-cell mouse embryos were cultured with either the dry or humid systems. Moreover after obtaining
informed consent 32 4-8 cell human embryos were also randomly assigned to culture with one of the above
systems. All media droplets were covered with mineral oil and the media were exchanged every 48 hours.
Blastocyst development rates were evaluated after 3 days. Results: For mouse embryos 93.1% and 91.9%
developed to blastocyst stage with the dry and the humidfied incubator system, respectively. Similarly theblastocyst formation rates of human embryos used in the experiment were not significantly different 62.5% and56.3% with the dry and humid incubator, respectively. Conclusions: Our results suggest that the development ofboth mouse and human embryos in a dry culture system were effectively identical to those of embryos cultured
under standard humidified culture conditions. Although results are still preliminary regarding human application in
our opinion, it might not be necessary to use a humidified atmosphere for culturing mouse or human embryos. The
dry culture system used in the present study is efficient, hygienic and requires only little maintenance. Hence thisnew dry incubation system could reduce the risk of a possible contamination and contribute to lowering
maintenance costs of an in-vitro fertilization program.

P-070

Contribution of oocyte and embryo cleavage quality in obtaining blastocysts
1

1

1

2

2

○Mounir Ajina , Sonia Jallad , Souhir Mehri , Sawsen Meddeb , Hedi Khairi , Ali Saad1

3

2

Unit of reproductive Medicine. University Hospital Farhat.Hached, Sousse, Tunisia, Department of Obstetrics and Gynaecology,
University hospital Farhat.Hached, Sousse, Tunisia., 3Laboratories of Cytogenetic, Molecular Biology and Human Biology of
Reproduction Farhat Hached Hospital, Sousse-Tunisia

Introduction: Oocyte growth and quality are dependent on the normal growth and differentiation of the oocyte
harboring follicule. Oocyte quality is assessed by the absolute number of mature oocytes and number of embryos
used. The evaluation of Oocyte quality correlated with the rate of fertilization and embryo quality after ICSI. The
aim of this study was to assessing oocyte and cleavage embryonic quality according to the quality of blastocysts.
Material and Methods: This retrospective study included forty nine infertile couples, who undergo ICSI withtransfer of blastocyst at the unit of Reproductive Medicine of the University hospital Farhat Hached, Sousse,
Tunisia. Embryo transfer performed during 2009-2010. The average age of women and men were equal
respectively 32 ± 3.5 and 37.69 ± 4.92 years with duration of infertility equal 6.6 ± 3.68. Sperm and oocytes
preparation techniques for intracytoplasmic sperm injection and evaluation of embryo quality were interpreted
according to WHO criteria. Results: The mean number of aspirated oocytes was 9.3 ± 3.69. The majority ofoocytes were good qualities (65.3). Mean fertilization rate was 76.23 ± 18.1. The average percentage of abnormal
sperm forms was proportional to the average rate of blastocyst tops (p = 0.03). The average number of oocytes
obtained was proportional to the average number of blastocysts. The average number of cleavage embryos obtained(5.59 ± 2.38) was inversely correlated with the average number of blastocysts (1.51 ± 1.06). The oocyte maturationand fertilization rate were inversely proportional to the rate of blastocysts. The mean pregnancy rate was 44.9 %.
Conclusions: A high percentage of good qualities oocytes associated with high rates of fertilization were favorable
for obtaining blastocysts and good pregnancy rate. The average percentage of abnormal sperm forms was
proportional to the average rate of blastocyst tops (p = 0.03). But oocyte maturation and fertilization rate wereinversely correlated to the blastocysts rate.

295

●Poster Presentations

P-071

Involvement of first cytokinesis on gene expression at blastocyst stage○Satoshi Sugimura, Tadayuki Yamanouchi, Yutaka Hashiyada, Eiji Kobayashi, Kei ImaiNational Livestock Breeding Center, Fukushima, Japan

Although we showed that cytokinesis at first cell cycle affect post-transfer viability, the mechanism has been
unclear. The aim of present study is to investigate effect of duration and cleavage pattern at first cytokinesis on
gene expression at blastocyst stage that related to implantation and pregnancy reorganization. Bovine in vitro
fertilized (IVF) embryos were cultured in well of the well culture dishes contained 125 µl CR1aa medium
supplemented with 5% calf serum for 168 h post insemination (hpi). Development of the embryos was monitored
using time-lapse cinematography and embryos developed to blastocyst stage were individually examined gene
expression (AKR1B1, CDX2, IFNτ, IGF2R and PLAC8) using real-time PCR. As result in multiple regression
analysis, duration at first cell cycle had a correlation with IFNτ (P = 0.002) and IGF2R expressions (P = 0.005), and
IGF2R expression in embryos cleaved after 27.25 hpi was lower than in vivo derived embryos (P < 0.05). IFNτ
expression was also affected a number of blastomere (P = 0.014), and the expression level in embryos undergoingdirect cleavage from one-cell stage to three-four blastomeres was lower than those characterized by two
blastomeres and in vivo derived embryos (P < 0.05). Furthermore, AKR1B1 expression in embryos withfragmentation was higher than embryos without fragmentation and derived in vivo (P < 0.05). Higher pregnancy
rate observed for animals in which transferred blastocysts had undergone first cytokinesis within 27 hpi andindentified by the presence of two blastomeres without fragmentation, than among those with abnormal
cytokinesis. These results indicate that duration and cleavage pattern at first cytokinesis may involve to gene
expression at blastocysts stage, which are related to implantation and pregnancy reorganization, resulting in impact
of post-transfer viability. This work was supported by the Research and Developmental Program for New
Bio-Industry Initiatives.

P-072

Zona-free oocyte formed a one pronucleus zygote following piezo intracytoplasmic
sperm injection can subsequently develop to the blastocyst in micro-well culture
system. (Case Report)○Xinzhi Yang, Akina Takamura, Emi Fukunaga, Tomoyo Kusuda, Shinichiro Okano,
Masayuki Kinutani
Kinutani Women's Clinic, Hiroshima, Japan

Introduction: Before performing intracytoplasmic sperm injection (ICSI), cumulus-oocyte complexes are treated byhyaluronidase to remove their cumulus cells. Occasionally, the ooplasms can escape from their zona pellucida
during this procedure. The mature and morphologically normal zona-free oocytes can be fertilized with ICSI andthen cultured to the blastocyst stage (J. Ding et al. Human Reproduction, 1999). ,Material & Methods: A 33-year-old primary unexplained sterility woman underwent an ICSI cycle in our clinic.
The clomiphene citrate/HMG method was used in her ovary stimulation. Briefly, from day 3 of her menstruation,
100 mg clomiphene (Shionogi, Japan) was given every day for 7 days. From the day 7 of her menstruation, 150 IU
HMG (Fujipharma, Japan) was injected per 2 days for twice. On the day 9, 0.25 mg antagonist (MSD, Japan) and10000 IU hCG (ASKA, Japan) were injected. Finally, one oocyte-cumulus complex was retrieved on day 11. After
4 hours of culture in HTF medium (Irvine, CA) with 10% Serum Substitute Supplement (SSS) (Irvine Scientific,
CA) in 6% CO2 in air, the cumulus cells were removed by gently peppitting in 40 IU hyaluronidase in order toperforming ICSI. Unfortunately, the matured ooplasm was loosing its zona pellucida. The zona-free oocyte wasinjected with one spermatozoon from her husband by using piezo-ICSI and cultured in HTF medium with 10% SSS
in 6% CO2 6% O2 and 90% N2. After 16 hours of culture, the fertilized oocyte developed a one pronucleus (1PN)
zygote. The 1PN zygote was cultured in 50μl SICM medium with 10% SSS (COOK, Australia) in a micro-welldrilled by melting the 35 mm dish bottom with a heated steel rod. On the third day of culture, SICM medium wascarefully removed and 50μl SIBM medium (COOK, Australia) supplemented with 10% SSS was added tocontinuously culture. On the fifth day, no blastocyst was formed but a good expanding blastocyst was formed on
the eighth day. ,Conclusions: The results suggest that zona-free oocyte can be fertilized by piezo ICSI, and the fertilized zygote candevelop to a good blastocyst using micro-well culture system. Combine with the previous result that 1PN-derived
blastocyst could result in pregnancy and birth in human assisted reproductive technology (Lyn Gras et al., Human
Reproduction, 1999), the blastocyst derived from zona-free 1PN zygote may be used for clinical treatment.

296

●Poster Presentations

P-073

A CUMULATIVE EMBRYO SCORING SYSTEM USED TO PREDICTPREGNANCY OUTCOME○Yoon Jeong Choi, Jung Ho Kim, Chan Park, Kwang Rae Kim, Sung il Roh, Hee Jun chi
i-Dream Center, Mizmedi Hospital, Seoul, Korea

Objective: To investigate the efficiency of our Cumulative Embryo Score (CES) system for selecting the optimalembryos to transfer Design: A prospective study Materials and Methods: This study was performed on 436 IVF-ET
cycles from January 2010 to February 2011 in which one(64 cycles), two(206 cycles) or three embryos(166 cycles)
were transferred. The quality of embryos was assessed using our CES system. Early-cleavage embryos received
two points, but non early-cleavage embryos did not receive any point. In addition, we also gave the embryos apoint from 5 to 1 according to the grades of embryos (G1 to G5) on day 2 and 3, respectively. Therefore, eachindividual embryo could receive the point from 2 to 12. The CES of individual embryo was used to evaluate thedevelopmental potential of the embryos. The IVF-ET cycles were categorized into nine groups according to the
CES and the number of embryos transferred: CES group A1(score 9-12, 1ET), A2(score 19-24, 2ET), andA3(score 26-36, 3ET), CES group B1(score 5-8, 1ET), B2(score 13-18, 2ET), and B3(score 15-25, 3ET), CES
group C1(score 1-4, 1 ET), C2(score 1-12, 2ET), and C3(score 1-14, 3ET), respectively. Results: When one
embryo was transferred, the pregnancy rates of A1, B1 and C1 groups were 50.0%, 19.3% and 9.5%. When twoembryos were transferred, the rates of A2, B2 and C2 groups were 67.3%, 52.8% and 45.0%. When three embryos
were transferred, the rate of the A3 group was 72.0%, those of the B3 and C3 groups were 50.0% and 33.3%,
respectively. The mean CES/embryo in the pregnant group was 7.78 that were significantly higher than that in thenon pregnant group 6.23. When one embryo transferred, the mean CES/embryo in the pregnant group was 7.93, but
the CES in the non pregnant group was 5.54(P<0.05). When two embryos transferred, the mean CES/embryo in the
pregnant group was 7.85, but the CES in the non pregnant group was 7.00. When there embryos transferred, themean CES/embryo in the pregnant group was 6.93, but the CES in the non pregnant group was 6.72.Conclusion: In
the present study, a close correlation between the pregnancy rates and the CES was observed. The cut-off values of
the CES/embryo according to the number of embryos transferred in the pregnant groups may be a good indicator to
select the embryo and to decide the number of embryos for transfer.

P-074

Cytoplasmic halo as an additional indicator to predict the quality of the results of
IVF and ICSI embryos○Devi Natalia, Harris Harlianto, Sintya Jatnikasari, Tono Djuwantono, Wiryawan Permadi
Aster Clinic, RS.Hasan Sadikin, Bandung, Indonesia

Introduction: The important thing that determines the success of IVF is selection the best viable embryos
transfered.There are some assessment such as Z scoring system,fragmentation and number of
blastomere.Sometimes, we look the clear area on the cortex of zygote at the day of fertilization check, calledcytoplasmic halo. It is due to the movement of cytoplasmic organelles specially mitochondria to the center, closer
to the pronucleus. Several previous studies mentioned, the formation of cytoplasmic halo at zygote stage is theindication of good quality embryos (Gada,et al,2007;Shimura,et al,2006). But these studies have not compared
between ICSI and IVF. So, in this study, we compared the number of embryos that presence halo and the quality ofembryos resulted between ICSI and IVF. Material & Methods: The study was conducted on embryos of 55 Aster
Clinic patient in January 2010 to May 2011(34 ICSI and 21 IVF). At zygote stage, along with fertilization check,the presence of the halo was recorded. On the day of ET, embryo quality was assessed (excellent, good, moderate,poor) based on number of blastomeres, fragmentation, blastomere size and suitability. Result: We found that both
fertilization program (IVF and ICSI), percentage of excellent and good embryos (23,15% and 48,05%) was
significantly higher on embryos halo(+) than embryos halo(-)(19,67% and 8,95%). While the percentage ofmoderate and poor embryos on embryos halo (+)(19,67% and 8,95%) lower than embryos halo (-) (30,27% and17,04%). The number of zygote halo (+) in ICSI (54,56%) is significantly lower than IVF (75,38%). While thepercentage of zygote halo (-) in ICSI (46,02%) is significantly higher than in IVF (24,71%). This is also supportedby the lower quality of embryos performed ICSI than IVF, either on embryos that have a halo or not. In ICSI, thepercentage of excellent, good, and moderate embryos was lower than IVF embryos, while the percentage of poor
quality embryos was higher. The lower embryo quality in ICSI embryos probably caused by more treatment to the
oocytes prior to ICSI performed. Its damage caused the migration of mitochondria in ooplasma disturbed so thehalo was not formed and the lack of ATP around pronucleus which produces the low quality embryos (Fujimoto,etal,2011). Conclution: Cytoplasmic halo can be an additional indicator for predicting the embryo quality fertilized
by ICSI and IVF. Percentage of embryos with halo (+) was higher in embryos fertilized by IVF compared to ICSI.

297

●Poster Presentations

P-075

Withdrawn

P-076

Effects of L/D stimuli and the circadian clock on reproductive physiology in female
mice
1

2

○Tomoko Amano , Juergen Ripperger , Urs Albrecht
1

2

2

Kinki University, Japan, Dep. of Medicine, Div. of Biochemistry, University of Fribourg

Background: The effects of light/dark (L/D) and the circadian clock on physiology should be separately examinedbecause L/D stimuli influence not only physiology but also the circadian clock. Results: We uncoupled the two
factors by comparing the reproductive physiology including estrous cycle progression and ovulation profile ofwild-type mice and that of mice that had completely lost the circadian clock, Per1/Per2 double-knockout(Per1/Per2) mice and Per2/Cry1 double-knockout (Per2/Cry1) mice, under L/D and constant darkness (D/D).
Under L/D, no abnormality was observed in estrous cycle progression, but perturbation of timely ovulation andaccompanying decline of reproductive ability were observed in Per1/Per2 and Per2/Cry1 mice. Although
pituitary-derived LH surge, which is prerequisite for ovulation, occurred at almost the same timing in wild-typemice and Per1/Per2 mice, expression pattern of Cox2, the gene responsible for timely ovulation, was perturbed in
the ovary of Per1/Per2 mice. Since overexpression of CLOCK and BMAL1 and REV-ERB ALPHA, componets ofthe circadian clock, suppressed the expression of Cox2 in cultured ovarian granulosa cells, respectively, Cox2
expression is linked to the circadian clock. Taken together, the data suggest that the perturbation of timely
ovulation in Per1/Per2 mice was likely due to the lack of the ovarian circadian clock. Under D/D, although noabnormality was observed in estrous cycle progression in wild-type mice, Per1/Per2 and Per2/Cry1 mice showed
drastic elongation of estrous stage, which caused elongation of the whole estrous cycle. Both wild-type mice andPer1/Per2 mice showed abnormal ovulation profiles as observed in Per1/Per2 mice kept under L/D. Conclusions:
Our results indicate that cooperation of L/D stimuli and the circadian clock optimizes female reproductive
performance.

298

●Poster Presentations

P-077

Numbers of CGG repeats on the FMR1 gene in Japanese patients with premature
ovarian insufficiency○Naoki Okamoto, Naomi Hamada, Yodo Sugishita, Nobuhito Yoshioka, Noriyuki Takahashi,
Bunpei Ishizuka
Department of Obstetrics and Gynecology, St.Marianna University School of Medicine, Kanagawa, Japan

Objective: To define the number of CGG repeats on FMR1 in Japanese patients with premature ovarian
insufficiency (POI) and normal controls and to identify correlations between number of CGG repeats and age at theonset of amenorrhea in these patients. Design: Retrospective, controlled cohort study.Setting: Outpatient
department of academic tertiary center. Patients: One hundred and twenty-eight consecutive Japanese patients withsporadic, non-syndromic POI and 98 controls with normal menstruation.Interventions: DNA was obtained fromplasma of each subject. Main Outcome measures: Whether the distribution of numbers of CGG repeats differs
between patients with POI and controls and whether the number of these repeats correlates with age at the onset of
amenorrhea in such patients.Results: We identified ≥40 repeats in all alleles except for eight in seven patients with
POI with intermediate numbers of repeats and two in the premutation range in two patients. The prevalence of >40
repeats was significantly greater in the patients. Age at the onset of amenorrhea negatively correlated with ≥36
CGG repeats in patients with POI. Conclusions: Over 36 CGG repeats in the FMR1 might intensify the etiology of
POI at least up to the premutation range.

P-078

Genetic analysis of spermatozoa in four infertile patients with macrocephalicsperm head syndrome
1,2

3

4

1

1

○Ghaya Merdassi , Pierre Ray , Meriem Chaabouni , Nedia Memmi , Fethi Zhioua , Amel Zhioua
1

2

1

3

IVF Center, Aziza Othmana Hopsital, Tunisia, Faculty of pharmacy.Monastir.Tunisia, Department of genetic and procreaction.
Tronche Hospital .grenoble. France , 4Department of human genetic. Charles Nicolle Hospital .Tunisia

Objective: Total teratospermia with predominantly macrocephalic (large headed) spermatozoa is a rare
abnormality. The purpose of this study was to evaluate the chromosomal content of spermatozoa in cases ofmacrocephalic sperm head syndrome (SM) and to describe the association between various phenotypes, sperm
chromosomal abnormalities, and the mutation of the aurora kinase C gene (AURKC). Materiels and methods: The
patients were four infertile patients with macrocephalic sperm head syndrome (91%, 82%, 100% and60%respectively). Morphological aspects for MS were detailed after shorr staining. The chromosomal content of
the spermatozoa was analyzed by fluorescent in situ hybridization (FISH) using X, Y, 18 centromeric probes.
AURKC sequence analysis was carried out for all patients. Result(s): Two morphological aspects for MS werehighlighted: MS with irregular head and multiple flagella, MS with regular head, all the nonselected spermatozoa
were abnormal, diploid, or polyploid. Sequence analysis showed that the 3patients with total syndrome were
homozygous for the AURKC c.144delC deletion. Conclusion(s): In conclusion, we confirmed that the absence of
AURKC c.144delC is not a sufficient condition to indicate ICSI in cases of SM. In fact, the very low proportion of
normal haploid spermatozoa contraindicated ICSI for our four patients. We suggest performance of these analysis
before including (or not), in an ICSI program, patients with macrocephalic sperm head syndrome.

299

●Poster Presentations

P-079

Gene Expression Profiling of Karyotypically Normal, Trisomy 12 and XXY Human
Embryonic Stem Cells
1

1,2

2

2

1,2

Hye Won Seol , ○Sun Kyung Oh , Baik Seol Cho , Kyung Eui Park , Young Min Choi ,1,2Shin Yong Moon
1

Institute of Reproductive Medicine Population, Medical Research Center, Seoul National University College of Medicine,
Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul 110-799, Korea

2

Human embryonic stem cells (hESCs) are well known to have the ability to proliferate unlimitedly and todifferentiate into all cell types. Recently, a number of reports have been published in which chromosomal
abnormalities has been observed in hESCs. We analyzed gene expression profiles of normal cell lines (SNUhES3:
S3 and SNUhES4: S4), two variants of SNUhES3 (trisomy 12: S3v12 and XXY: S3vX), and one variant of
SNUhES4 (trisomy 12: S4v12) using the Illumina HumanRef-6 BeadChip including 48,095 probe sets to findepigenetic difference between normal cells and chromosome variants. Gene expression patterns were showed that55 genes were up-regulated and 28 genes were down-regulated in S3v12 to compare with S3 and 98 up-regulated
genes and 75 down-regulated genes were identified in S3vX. Also, 87 genes were up-regulated and 61 genes were
down-regulated in S4v12 to compare with S4. In particularly, three genes, PUS7L (synthetase), INHBE (growth
factor) and PKIB (kinase inhibitor), were up-regulated and two genes, ERG1 (KRAB box transcription factor) and
GALNT9 (glycosyltransferase) were down-regulated in S3v12, S4v12 and S3vX compared to normal hESC lines.
Gene expression profiling in variant hESC lines with trisomy 12 or XXY chromosome as well as normal hESC
lines may provide insights to the development of research field for cell differentiation and clinical approach such ascell replacement therapy. This research was supported by a grant (SC-1150) from Stem Cell Research Center of the
21st Century Frontier Research Program funded by the Ministry of Education, Science and Technology, Republicof Korea.

P-080

Effect of dibutyryl cAMP during in vitro maturation on the developmental
competence of bovine oocytes after ICSI
1,2

2

2

2

○Chikako Kani , Tomohisa Wada , Akiko Kuwahata , Masanori Ochi , Toshitaka Horiuchi
1

1

2

Graduate School of Comprehensive Scientific Research, Prefectural University of Hiroshima, Hiroshima 727-0023, Japan, Ochi Yume
Clinic Nagoya, Aichi, Japan

Introduction: Bovine intracytoplasmic sperm injection (ICSI) is useful technique for production of calves fromlimited supply of gametes. In bovine ICSI, in vitro-matured oocytes are used, but it is well known that thedevelopmental competence of in vitro-matured oocytes is lower than that of in vivo-matured oocytes. The content
of cAMP of follicular oocytes is increased during follicular development, and differ from individual immatured
oocytes. The objective of this study was to examine effect of treating bovine oocytes with dibutyryl cAMP
(dbcAMP) during in vitro maturation (IVM) on nuclear maturation and embryonic development after ICSI.
Material& Methods: As a basic medium, TCM199 supplemented with 1mg/ml recombinant human albumin, 1
IU/ml recombinant human FSH and 50 ng/ml EGF was used. In the first experiment, bovine cumulus-oocyte
complexes (COCs) were cultured in the basic medium supplemented with 0, 10, 100 and 1000 μM dbcAMP for 21
h. We examined the effect of the concentration of dbcAMP during IVM on nuclear maturation and the blastocyst
development after ICSI. In the second experiment, we examined the effect of treating COCs with 10 μM dbcAMP
on sperm aster formation in bovine IVM oocytes at 5 h after ICSI. In the third experiment, using time-lapse
microscopy, we observed the effect of treating COCs with 10 μM dbcAMP during IVM on the speed of embryo
development after ICSI. Results: In the first experiment, treating COCs with 0 to 1000 μM dbcAMP during IVM
did not affect the percentage of MII oocytes (69 to 79 %). Blastocyst rate in the group of 10 μM dbcAMP (56%)
was significantly higher than that in the others (0 μM: 24%, 100 μM: 43% and 1000 μM: 31%). In the second
experiment, the percentage of sperm aster formation in bovine IVM oocytes treated with 10 μM dbcAMP was
significantly higher than that in the untreated IVM oocytes (82 % vs. 52%). In the third experiment, timing ofcleavage and blastocyst development after ICSI in the group of 10 μM dbcAMP was significantly faster than that in
the untreated group (cleavage, 22.2±0.5 h vs. 25.6±0.8 h, blastocyst, 162.1±2.2 h vs.170.3±3.2). Conclusions: We
demonstrate that treating bovine COCs with 10 μM dbcAMP together with FSH and EGF during IVM does notaffect nuclear maturation at 21 h, but stimulates the sperm aster formation after ICSI, accelerates the speed ofembryo development, and improves the blastocyst rate.

300

●Poster Presentations

P-081

Androstenedione induces abnormalities in morphology and function of developing
oocytes, which impairs oocyte meiotic competence
1

1

1

1

2

○Wataru Tarumi , Sanae Tsukamoto , Yuki Okutsu , Noriyuki Takahashi , Masanori Itoh ,1
Bunpei Ishizuka
1

Department of Obstetrics and Gynecology, St. Marianna University School of Medicine, Kanagawa, Japan., 2Department of Biology,
College of Liberal Arts and Sciences, Tokyo Medical and Dental University, Ichikawa, Chiba 272-0827, Japan.

Previous studies have suggested that intraovarian excess androgen, including androstenedione, is a main cause ofpolycystic ovary syndrome. However, the mechanism leading to polycystic ovary by androgen is currently unclear.
To obtain further insight into the effects of androgen on ovarian folliculogenesis and oogenesis, early secondary
follicles were isolated from the mouse ovaries and were cultured individually in vitro with or without
androstenedione (10-11-10-5 M) for 12 days. In this single follicle culture system, early secondary follicles could
develop to preovulatory stage. Androstenedione treatment reduced survival rates of follicles and promoted theformation of follicles with abnormal morphology, such as the lack of cumulus and mural granulosa cells andmisshapen oocyte. Significantly higher estradiol secretion was observed in androstenedione-exposed follicles. In
addition, androstenedione treatment prevented the alteration in chromatin configuration and influenced expressionof growth differentiation factor 9 in the oocytes. When follicles cultured with androstenedione for 12 days weretreated with human chorionic gonadotropin and epidermal growth factor, the first polar body exclusion,
chromosome alignment on metaphase plate, and spindle assembly were inhibited in the oocytes. These results
demonstrate that excess androgen and perhaps its metabolite, estrogen, induce abnormalities in morphology andfunction of developing oocytes, which impairs oocyte meiotic competence.

P-082

Preimplantation genetic diagnosis for spinal muscular atrophy usingmini-sequencing and genetic linkage analyses at the blastocyst stage
1,2

3

3,4

○Chia-Cheng Hung , Shee-Uan Chen , Shin-Yu Lin , Yi-Ning Su

1,2,4

1

Graduate Institute of Clinical Genomics, National Taiwan University College of Medicine, Taipei, Taiwan 2, Department of Medical
Genetics, National Taiwan University Hospital, Taipei, Taiwan, 3Department of Obstetrics and Gynecology, National Taiwan
University Hospital, Taipei, Taiwan, 4Graduate Institute of Clinical Medicine, National Taiwan University College of Medicine, Taipei,
TaiwanBackground: Approximately 94% of spinal muscular atrophy (SMA) cases are caused by common homozygous
absence of the SMN1 gene. Although preimplantation genetic diagnosis (PGD) is an alternative option for couples
at risk of having a child with single gene disorder, blastomere biopsy has been employed in most in vitro
fertilization procedures. Methods: In the present study, we validated and applied protocol clinically for PGD
through the use of blastocyst biopsy, whole genome amplification, mini-sequencing genotype coupling withgenetic linkage of SMN gene involving three informative microsatellite markers, and thawed embryo transfer.
Results: We report data to identify the SMN1 gene deletion on eighteen clinical embryos obtained from one
participating couple, where both partners are heterozygous SMA carriers with 1-SMN1/3-SMN2 genotypes.
Approximately 78% (14/18) of blastocysts were successfully amplified in a single PGD cycle. Among theseembryos, ten (72%, 10/14) were diagnosed as unaffected, two (14%, 2/14) as affected, and two embryos (14%,2/14) had no conclusive diagnosis due to allele drop-out (ADO). Two unaffected embryos were thawed andtransferred in the next cycle resulting in a singleton pregnancy, and the birth of a healthy girl who carries the
1-SMN1/3-SMN2 genotype. Conclusions: The strategy of PGD using blastocyst biopsy and thawed embryotransfer increased the reliability of the results and permitted more time for the execution of molecular diagnosis.
The improved protocol for PGD could be adopted for other monogenic diseases in the future.

301

●Poster Presentations

P-083

Preimplantation Genetic Diagnosis (PGD) for translocation carrier with mild
stimulation and single vitrified blastocyst transfer : Case Report○Naoki Aoyama,Yuji Takehara, Satoshi Kawachiya,Tomoko Kuroda, Nami Kawasaki,
Rie Yamadera,Keiichi Kato, Osamu Kato
Kato Ladies Clinic, Tokyo, Japan

Introduction: In population of chromosomal translocation carriers, hyperstimulation is still applied for PGD cycle to increase thenumber of retrieved oocytes, because it is considered that they have high risk of meiotic non-disjunction. Then, how many oocytes
does carrier couple need to conceive a healthy baby? This is the new approach for PGD with mild stimulation which is developed
from our experience in two decades. Material and methods: Twelve couples consisting of subjects with a mean age of 35.2 years
who are carriers of chromosomal translocations.The patients underwent mild ovarian stimulation, which was carried out by
administration of clomiphene citrate in combination with a minimum amount of urinary HMG (75IU X less than 4 times) or rec.
FSH (75IU X less than 3 times). Administration of clomiphene citrate was initiated from day 3 of the menstrual cycle at 50mg/day
and was continued until the day before administration of the GnRH agonist as the maturation trigger (GnRHa nasal spray, 300µg)
and 32-35 h before oocyte retrieval as described by Kato. IVF was performed for all oocytes, and blastomere biopsy was
performed on the embryos reaching at least the 6-cell stage on day 3. Culture of all the biopsied embryos was continued, andembryos developing to the blastocyst stage were vitrified. FISH analysis was performed on all biopsied blastomeres using 3 to 4
appropriate FISH probes specific for translocated segments. Cryopreserved blastocysts which showed normal/balanced were
thawed and intrauterine transfer of single blastocysts was performed. Results: Eighty-nine Cumulus Oocyte Complex (COC) werecollected in 35 OR cycles from 12 patients, average number of COC per OR cycle were 2.5(89/35). Ninety one % (77/85) werefertilized, 28% (20/72) were transferable at the 8-cell stage and 13 of the blastocysts which showed alternate were thawed andintrauterine transfer was performed by single blastocyst transfer for 8 patients. A positive heart beat was obtained in 7 patients,
pregnancy rate per ET was 54% (7/13). Finally, the delivery rate was 46% per ET (6/13) and 17% per OR (6/35), and one case isongoing at 20 weeks. There were 6 deliveries of 6 healthy babies, consisting of 2 males and 4 females, with no major
malformations at birth. Conclusions: ESHRE PGD consortium‘s data (2009) with 417 patients showed that the average numbersof COC to obtain a fetus were 64.1(No. of COCs/No. of FHB) which was five times of our data (12.7: 89/7). There haven‘t been
any reports concerning to assess the appropriate number of oocyte to obtain a pregnancy for carrier couple, however, our results
and subsequent natural pregnancy outcomes which was reported by Sugiura-Ogasawara may indicate that it is enough around 10
oocytes for a delivery. And this conclusion strongly suggests that to decrease the number of oocytes requires not only accurate
diagnosis skill but also improved IVF-ET technique in facilities.

P-084

Noninvasive morphological examination for germinal vesicle oocytes predict theirmaturity process of completion
1

1

1

1

1

○Tsuyoshi Okubo , Ryoko Matsuo , Naoko Shimada , Teruaki Hayashi , Tomoya Segawa ,1
2
Shoukichi Teramoto , Masashige Kuwayama
1

Shimbashi Yume Clinic, Tokyo, Japan, 2Repro-Support Medical Research Center, Tokyo, Japan

Objective: Especially, derived from small size follicle, germinal vesicle (GV) oocyte sometimes can be obtained in human IVF.
Those human GV oocytes have been researched as in vitro maturation (IVM) for a long time. However, IVM studies conductedwith GV stage oocytes have not produced promising results with regard to oocyte maturity and blastocyst development. We
noninvasively examined GV stage oocyte by means of morphology with microscope and analyzed whether or not thesemorphologic parameters can predict their maturity and quality. Method: This study was designed 197 IVF cycles from May 2010
to March in 2011. All GV oocytes were obtained from small size follicles (＜ 8mm diameter) for natural cycle IVF and cultured
in vitro in P1 medium (Irvine Scientific, USA). Cumulus cells were gently removed by pipetting with hyaluronidase, thencumulus-free GV oocytes were morphologically examined by contrast-phase microscope (Olympus Optical Co., Japan) at ×400
magnification. We measured two times in a different orientation and calculated the average diameter of whole cytoplasm, GV andchromatin configuration using by Octax Eyeware software (OCTAX Microscience GmbH, Germany). Cumulus-free GV oocytes
were cultured in P1 culture medium at 37℃ and 5% CO2, 5% O2 and 90% N2. All oocytes were confirmed the presence of GV or
first polar body and classified as immature (GV, MI) or mature (MII) after 24 hours in vitro culture. Results: A total of 198 GV
oocytes were obtained from 127 patients. Patient age varied from 24 to 48 years old (38.9 ± 4.0 years old). The GV oocytes wereretrospectively examined after removing cumulus cells. Of 148 GV oocytes were cultured for 24 hours, it was found that there
were 44 GV, 25 MI and 79 MII. (53.4% matured) Mean diameter of oocyte cytoplasm of GV, MI and MII were 114.75μm,
116.2μm and 116.44μm,respectively. The mean diameter of GV were 32.67μm, 32.37μm and 32.73μm, respectively. The mean
diameter of chromatin configuration were 8.54μm, 8.63μm and 9.06μm, respectively. In addition, 1.4% of GV oocyte contained
two chromatin configurations (2/148), these two oocytes remained in GV after 24 hours culture. Student-T test was used forstatistical analysis. Conclusion: Matured oocytes contained significantly larger size chromatin configuration than immature
oocytes (P<0.05). Different size of chromatin configuration may be related to GVBD and oocyte maturity. The oocyte which had
two chromatin configurations did not mature, therefore it appears that the morphology of chromatin configuration is relating
mechanism of oocyte maturation. Furthermore, specific morphologic parameters may provide information of oocyte maturation in
IVM..

302

●Poster Presentations

P-085

hCG is critical for the uterine decidual response in mice
1

1

1

1

2

○Hiroomi Kawano , Kenji Ezoe , Noriko Kagawa , Akiko Yabuuchi , Keiko Ochiai ,, Hiroshi
2
1
1
1
1
Nagashima ,, Hisao Osada , Fumihito Aono ,Yuji Takehara , Osamu Kato
1

Kato Ladies Clinic, Tokyo, Japan 2Meiji University, Kanagawa, Japan

Introduction: The process of implantation, necessary for all viviparous birth, consists of tightly regulated events, including
apposition of blastocyst, attachment to the uterine lumen, and differentiation of the uterine stroma. In rodents and primates
the uterine stroma undergoes a process called decidualization. Decidualization, the process by which the uterine endometrial
stroma proliferates and differentiates into large epithelioid decidual cells, is clitical to the establishment of fetal-material
communication and the progression of implantation.On the other hand, it was said that fertilization rate of hCG injected
mice was lower than normal mice. So, this our study investigate into the relation between decidualize after hCG injection inmice and their fertility. Material & methods: Mice were maintained in the designated animal care facility at Meiji
University according to the institutional guidelines for the care and animals. The artificial decidual response has beenpreviously described (Finn, C.A., et al., 1972, Biol. Reprod. 7:82-86). Briefly, mated female ICR mice (8-10 weeks old) by
castrated male after PMSG and/or hCG treatment (1time and/or 3times) were injected 25μl sesame oil into uterine horn at
3.5d.p.c. to induce decidualization. The contralateral horn was not traumatized and served as a control. These mice weresacrificed and extracted both uterine horns at 7.5d.p.c. The ratio of wet weight in uterine horn after sesame oil injection tothe other no-stimulated one was calculated. At the time of dissection, uterine tissues and ovaries were fixed with 4%paraformaldehyde (PFA). To perform histological analysis and count number of oocyte, these samples were thendehydrated by ethanol washes, embedded with paraffin, sectioned longitudinally and stained with hematoxylin and eosin.
Finally we perfomed embryo transfer to hCG injected mice to check the fecundability. Results: Whereas uterine horn ofcontrol group was able to decidualize, uterine horns of hCG injection all groups were not able to perform decidualization
significantly (untreated group: 22.58±3.32g versus hCG injection group: PMS+ hCG 1.10±0.03g, h 2.44±1.0g, 3(PMS+hCG) 2.16±0.56g, 3hCG 3.21±0.77g; p>0.05). In pathological analysis, the cells of uterine endometrium in hCG injectedgroup was not hypertrophied and polygon. Pregnacy rate of hCG injected group (50%) was half of control group(100%) after embryo transfer and the number of pups in hCG group (56.2%) was significantly fewer than control group(23.3%). The body weight of these pups were lighter than control group (hCG group: 1.2±0.39g versus control: 1.55±0.25g)
significantly. Conclusions: Our study showed that hCG is critical for the uterine decidual response and infecundity in mice.
It was essential to make clear the mechanism between decidual hypoplasia with hCG and infertility in the further
investigation.

P-086

Cytogenetic study in early spontaneous abortion after IVF and ICSI○Satoshi Kawachiya, Yuji Takehara, Keiichi Kato, Hisao Osada, Naoki Aoyama, Osamu Kato
Kato Ladies Clinic, Tokyo, Japan

Objective: To investigate the incidence of chromosomal abnormalities in early pregnancy loss after IVF and ICSI.
Methods: From January 2004 to December 2007, a total of 166 cases of miscarriage villi samples after IVF-ET werecollected for cytogenetic analysis. The average age of cases was 38.9. All IVF cycles were performed by natural cycleor mini IVF in our center. Cytogenetic analysis was performed at special biochemical analysis center (SRL, Japan).
Results: Cytogenetic results were obtained for 158 (ICSI 63, IVF 95) samples and 117 samples (74.1%) showedchromosomal abnormalities. Autosomal trisomies of chromosomes 4, 7, 8, 9, 11, 12, 13, 14, 15, 16, 17, 18, 20, 21 and
22 were detected in 80 samples (68.4%). In 80 samples, 22 samples were trisomy 22(27.5%), 17 samples showedtrisomy 15 (18.8%) and 10 samples were trisomy 21 (12.5%). Double trisomy was detected in 13 samples. In 117
samples, 8 samples showed sex chromosome monosomy and all samples showed 45,X. Triploidy was detected in 1
sample and tetraploidy was detected in 3 sample. All of polyploidy abnormality were performed by conventional
insemination. Structual rearrangements were detected in 8 samples (6.8%). Normal karyotype samples accounted for
41 (25.9%). ICSI samples more showed aneuploidy than conventional insemination patients; 82.5% vs. 68.4%(P<0.05). There was no difference of average age between two groups. Comparison between conventional
insemination and ICSI did not show significant difference in the type of chromosome abnormalities. Conclusion:
Chromosome abnormalities are identified in 74.1% of all spontaneous abortions. In this study, the average age of IVF
patients is 39.5. Due to this background, incidence of abnormal chromosome of miscarriage villi showed higher thanprevious reports. In autosomal trisomy samples, incidence of trisomy 22 was particularly higher than otherchromosome trisomies. Furthermore, the frequency of trisomy 15 was higher than previous reports. These results
indicate that the proportion of chromosomal abnormalities may be different between normal pregnancy and IVF or
ICSI. We also found that ICSI samples were more likely to have aneuploidy than IVF samples.

303

●Poster Presentations

P-087

Day 7 -hCG level after frozen-thawed blastocyst transfer can be a predictor for
pregnancy outcomes?
1

2

1

1

1

○Keiichi Kato , Tomoya Segawa , Hisao Osada , Tamotsu Kobayashi , Yuji Takehara ,1
Osamu Kato
1

Kato Ladies Clinic, 2Shimbashi Yume Clinic, Tokyo, Japan

Introductions: There are only few large-scale studies about outcomes after single embryo transfer. In this time, we
analyzed the results after frozen-thawed single blastocyst transfer to find out the correlation between serum-hCG
levels supplied from single implanted embryo and pregnancy outcomes. Materials and methods: We investigatedtotal of 7,523 cycles of frozen-thawed single blastocyst transfer performed from Jan. 2004 to Dec. 2008. The age ofpatients was from 25 to 44 years. All cases were confirmed both serum -hCG levels on day 7 after blastocyst
transfer and the results about pregnancy outcomes. And also were excluded all multiple pregnancies and ectopic
pregnancies. All patients retrieved oocyte under the natural cycle or clomiphene based minimal stimulation
protocol. All obtained blastocyst vitrified using Cryotop® (Kitazato, Japan). Frozen-thawed single blastocyst
transfer performed on day 5 after spontaneous ovulation or on cycle day 18 in hormone replacement cycles. Serum
-hCG levels checked out 7 days after transferred embryo. Patients were divided into eight groups depending on
the -hCG levels. Pregnancy rates and live birth rates in each -hCG level were analyzed in 3 age groups (a. 25-34
year, b. 35-39 year, c. 40-44 year). Results: Patients with day 7 -hCG levels >30mIU/mL showed comparatively
high pregnancy rates and live birth rates regardless age group. Conclusions: Our study suggested that day 7 -hCG
level after single frozen blastocyst transfer may be used for a predictor for pregnancy outcomes.

304

AUTHOR INDEX

305

●Author Index – Special Guest Lecture, Pre-Congress Workshops, Plenary Lectures, Concurrent Symposia
P: Plenary Lecture; W-V: Pre-Congress Workshop - Vitrification
W-P: Pre-Congress Workshop – PGD; C: Concurrent SymposiumA

Feldberg

Dov

C-9.4

Friedler

Shevach

ISF 6

Frydman

Nelly

C-14.1C-16.1

AkutsuHidenori

C-23.1

FrydmanRené

STGO(1)2

Al Hasani

Safaa

C-8.3

Fujii

TakumaC-6.3

Allahbadia

Gautam

C-20.2

Fujiwara

HiroshiC-21.3Andersen

Claus

Yiding C-7.2

FukudaAisakuC-15.4Ando

MasaakiC-6.4

AoyamaNaoki

W-P. 4

G

Asada

Yoshimasa

C-19.1

García-Otero
Rufino

C-6.1

Ata

Baris

C-3.2

Reina

Azumaguchi

AtsushiC-16.3

Gomel

VictorOpening Ceremony
Opening LectureB

P-6
C-17.2

Gonen

Yael

C-21.1

Gürgan

Timur

C-2.4

Handyside

Alan

P-4

HashimotoShu

C-15.2

Hiramatsu

Yuji

C-5.2

Ho

Pak-ChungC-3.4

Benkhalifa

Moncef

C-16.2

Benkhalifa

Moncef

STGO(2)2

Ben-Rafael

Zion

C-3.8

BernardArtur

C-11.1

H

Boutaleb

Youssef

STGO(1)1

Brockerhoff

Peter

C-10.1

C-5.3

C
Cha

Kwang Yul

C-23.4

Huang

Guoning

C-11.2Chen

Hai Ying

C-14.6

Hurwitz

Arye

ISF 2

Chen

Hsin-Fu

C-19.4C-23.3Chian

Ri-Cheng

C-10.3C-14.4

Coll

Oriol

C-20.4

BunpeiC-19.3

D

C-4.4

C-22.4I
IshizukaJ

DeRosaMichaelC-5.5

JaffarMuchsin

C-1.3

Diedrich

Klaus

P-7

JanischClaus Peter

C-20.5

Dirnfeld

MarthaISF 3

Do

Byung-Rok

APART 1K

Dubuisson

Jean-Bernard

C-5.1

KagawaNorikoW-V.2

Katagiri

Yukiko

C-9.3

Kato

Osamu

P-1.1

F
Feichtinger

Wilfried

C-22.2

KovacsPeter

C-16.5Feki

Anis

C-23.2

KubotaToshiro

C-2.3

306

APART3

●Author Index – Special Guest Lecture, Pre-Congress Workshops, Plenary Lectures, Concurrent Symposia
Kuwayama

Masashige

W-V.4

C-8.2
Q

L

Qiao

Lebbi

Issam

C-4.2

Leibo

Stanley

W-V.1

Leong

MiltonC-13.2

Li

ShangweiC-20.1

Lichtenegger

Werner

C-6.2

Lim

Jin-HoC-15.3

Lindenberg

Svend

C-3.6

C-8.4

Jia-Yin

C-9.1

Lobo

Rogerio A.

C-1.2

Lunenfeld

Bruno

C-3.1

C-15.1Rabinovici

Yaron

ISF 1

Revel

Ariel

ISF 5

Luis Arturo

C-8.1

SallamHassan

C-3.7

Sato

SuguruW-P.2

SeidmanDanielC-20.3Seki

Moritoshi

C-21.4

Shibahara

Hiroaki

C-2.5

Shozu

Makio

C-2.1

SilberSherman

P-5

R

Ruvalcaba
Castellón
C-15.5C-16.4
Liu

Jie

S
C-4.1

C-7.4
ISF 4

M
Mahmoud

KhaledC-1.4

MakinoTsunehisa

C-12.1

Merdassi

Ghaya

STGO(2)3

Moon

Shin Yong

P-3

Mori

Takahide

C-1.1

Morimoto

Yoshiharu

C-14.5

MukaidaTetsunori

C-10.4

Murtinger

Maximilian

C-3.3

W-V.3

C-18.1
Smitz

Johan

C-13.3C-14.3

SueokaKou

W-P.1

C-22.3

Takakuwa

KouichiC-21.2

Takehara

Yuji

APART 5

Tan

Seang Lin

P-9

TanakaAtsushiC-17.3

TanakaAtsushiC-22.1

Teramoto

ShokichiC-7.1

TerrasKhaledSTGO(2)1

Tulusan

Liong

C-7.3

Tur-Kaspa

Ilan

W-P.3

Tzeng

Chii-Ruey

C-2.2

Vajta

Gábor

C-12.4

Vanderpoel

SherylC-20.6

Vanderzwalmen

PierreC-18.3T

N
Nabeshima

Hiroshi

C-4.3

Nagy

Zsolt Peter

P-2

NakamaKen

APART 2O
Ogawa

TakehikoC-23.5

Ombelet

Willem

C-20.7

Osada

Hisao

C-5.4

OumzianeAmina

STGO(1)3P
Palermo

Gianpiero D.

P-8

C-11.4

C-9.2

V

307

C-12.3

●Author Index – Special Guest Lecture, Pre-Congress Workshops, Plenary Lectures, Concurrent Symposia

W
WiwekoBudi

C-19.2

YabuuchiAkiko

C-17.1

Yamadera

Rie

APART 4

Yamanaka

ShinyaSpecial Guest
LectureYee

Bill

C-13.1Yoon

Tae-KiC-12.2

YoshidaHiroakiC-14.2

YoshidaAtsumiC-18.2

Zech

NicolasC-11.3Zech

Herbert

C-13.4

Zhang

John

P-1.2

ZhiouaFethi

C-10.2

Y

Z

C-3.5

308

●Author Index – Oral Communications and Poster Presentations
O: Oral Communication; P: Poster Presentation

BroomandA

C

FarzanehO-035

AbsalanForouzan

P-031

Cai

Li-Yi

P-007

Achache

Hanna

O-009

CASTELLO

DAMIA

P-052

AdachiTomokoO-033

Chaabouni

MeriemP-078

Aisaka

Kohzo

P-028

Check

Jerome H

P-006 P-009

Ajina

Mounir

P-064 P-070

P-012 P-022

AkhondiMohammad Mehdi

P-055

P-030 P-048

Alamolhoda

Zahra

P-053

Chen

Shee-Uan

P-082

Albrecht

Urs

P-076

Cheon

Kangwoo

P-060

Almog

Benny

P-063

Chi

Hee JunP-060 P-073

Amano

TomokoP-076

Chikazawa

Kenro

P-023

Amini mohabadi Javad

P-003

Cho

Baik Seol

P-079

Amo

Ami

O-024

Cho

J.D.

P-025 P-029

Angel

Marlane

P-068

Choe

Jung K

P-006 P-009

Angelova

Pepa. A

P-021

P-022 P-030

Ao

Lei

O-023

P-048

Aono

FumihitoP-033 P-069

Choi

Bum-Chae

P-016

P-085

Choi

Jin-Woo

P-015

AoyamaNaoki

P-083 P-086

Choi

Yoon Jeong

P-073

Arakawa

Osamu

P-054

Choi

Young Min

P-079

Araki

Yasuhisa

O-012

Cira

Nadir

P-068

Asada

Yoshimasa

P-008

CitrinoGabrielle

P-009 P-030

Askarian

Saeedeh

P-042

COBO

ANA

P-052

Cohen

RachaelP-012

CosteaGeorge

P-056

B
Baek

Kwang-Hyun

P-015 P-016

Bahadir

SelcenP-020 P-045

D

Bahadur

Anupama

O-038 P-061

De La Macorra

Angel

P-052

Baltadzhieva

Daniela. N

P-021

Deng

Bo

O-028 O-029

Barasila

Atikah

O-010

O-031 P-017

Barkn

Gali

P-063

P-044

Batioglu

A. Sertac

P-045 P-020

Deng

Lian

O-031 P-017

Beigi Boroujeni

MandanaP-013

DiAntonioAnn

P-009 P-012

Beigi Boroujeni

Masoud

P-013

Beigi Boroujeni

Nasim

P-013

Dimitriadis

E

O-032

Bhatt

Ashok

P-061

Dix

Ebony

P-012

Boukadida

Jalel

P-064

Djuwantono

Tono

P-074

P-022

309

●Author Index – Oral Communications and Poster Presentations
Dong

Ming-Yue

O-016

Hardiyanto

L.

O-001

Doosti

Rita

O-035

Harlianto

HarrisP-074

DouchiTsutomu

O-040 O-041

Hasegawa

A.

O-001

Hashiba

Tsuyoshi

P-058

Hashimoto

Hiromi

O-014

HashimotoShinako

P-062

Hashimoto

Shoko

P-043

Hashimoto

Shu

O-024

Hashiyada

YutakaP-071

Hashizume

RyoichiP-011

E
Ebrahimi

Bita

P-032

Eizenmann

Einat

O-025

Endo

Takumi

P-065

Endo

Yuji

P-047

EskandariMorad Pasha

P-003

Kenji

P-033 P-085

Nasab
Ezoe
F

Hassani Bafrani Hassan

P-003 P-004

Hatakeyama

Naohisa

O-026 O-027

Hayashi

MasaruO-036

Farrokhi

Ali

P-049

Hayashi

MasatoP-011

Forgacs

Vince

O-021

HayashiTeruaki

P-084

Forouzandeh-M Mehdi

P-010

Hestiantoro

Andon

O-010

Fujihara

Mayako

P-002

Hibi

Hatsuki

P-008

Fujii

JunichiO-022

HinokioKenji

P-018 P-067

Fujii

YoshitakaP-047

Hiraike

HarukoP-028

Fukunaga

Emi

P-072

Hiraike

Osamu

P-028

Fuse

Miki

P-065

Hirano

Yuki

P-023

Fuykasawa

Ichio

O-036

Hirayama

Kazuhiro

P-046

Fuzii

Osamu

P-005

Hirohama

JunO-026 O-027

Hiura

Rie

O-026 O-027

G

Hori

Sayumi

P-051

Gao

Mengying

O-031 P-017

HoriuchiToshitakaP-080

GhasemiMaryam

P-001

Hormozi

Maryam

P-055

Ghasemzadeh

Jalal

O-003

Hosseini

H

O-032

Ghorbanmehr

Nasim

P-053

Hosseinigohari

Ladan

P-055

Gibo

Akiko

O-040 O-041

Hu

han

P-027

Goel

SadeepP-002

Hu

Lin-Qing

P-007

Gu

Bon-Hee

P-016

Huang

Guoning

O-030

Gupta

Prerna

O-038

Huang

He-Feng

O-016

Gurlek

Beril

P-045

Huang

Weidong

O-002

Hung

Chia-Cheng

P-082

Hyodo

Hiroe

P-028

Hyodo

Hironobu

P-028

H
Hamada

Naomi

P-077

Hamatani

ToshioP-058

Hannachi

Neila

P-064

310

●Author Index – Oral Communications and Poster Presentations
I

Kani

Chikako

P-080

Iba

YumikoO-013 O-015

KaracaMujdegul

P-045 P-020

Ichikawa

Tomohiko

P-065

Karaca

Simla

P-045

Iida

SyuichiP-005 P-046

Karakida

Sinya

P-038

P-062

KataokaNobuhikoO-014 O-018

Kato

KeiichiP-035 P-033

Ikeda

Tomoe

P-023

Ikegami

Miki

P-019

P-083 P-086

Imai

Hiroshi

P-002

P-087

Imai

Kei

P-071

Imajo

Akifumi

O-013 O-015

P-033 P-083

ImamotoTakashi

P-065

P-085 P-086

IraharaMinoruP-018 P-067

P-087

Ishida

Eri

P-014

Katou

Masahiro

P-046

Ishihara

Osamu

O-020 P-036

Kawachiya

Satoshi

P-083 P-086

P-037

Kawamura

Toshihiko

O-040 O-041

Kato

Osamu

P-035 P-069

P-041

Ishizuka

Bunpei

P-077 P-081

Ito

KeijiroO-024

KawanoHiroomi

P-085

Ito

Masao

P-011

KawanoYasushi

P-038

Itoh

Masanori

P-081

Kawasaki

Nami

P-083

IwamotoMitsuru

P-062

Khairi

Hedi

P-070

IwashitaM

O-008 O-032

Kim

Giyoung

P-060

Iwata

Kyoko

O-013 O-015

kim

Jonghyun

P-060

Kim

Jung Ho

P-073

Kim

Kwang Rae

P-073

Kim

S.K.

P-025 P-029

Kim

Sung-Min

P-002

Kimura

Naoko

O-022

Kinutani

MasayukiP-072

Knaggs

Paul

O-006

Kobayashi

Eiji

P-071

Kobayashi

Tamotsu

P-087

Kobori

Yoshitomo

O-034

Kokeguchi

ShojiO-014 O-018

KomoriS

O-001

KosakaNobuaki

O-036

KoshidaMitsunobu

O-034

Kovalsky

Dina

P-063

KRAMP

F

P-052

J
Jallad

Sonia

P-070

Jamison

Theresa

P-006

Jatnikasari

SintyaP-074

Jeddi-Tehrani

Mahmood

P-055

JeoungHyo Young

P-015

Jinno

Masao

O-026 O-027

K
Kabir-Salmani

Maryam

O-008 O-032
P-039 P-059

KagawaNorikoP-033 P-085

Kai

Yoshiteru

O-015

Kajihara

Takeshi

P-036 P-037

Kameyama

Yuichi

P-011

KanasakiHaruhiko

P-057

311

●Author Index – Oral Communications and Poster Presentations
Kuang

YanPing

O-042

MalceaLenuta

P-056

Kudoh

Ken-ichiP-011

MalhotraNeena

O-038 P-061

Kuji

Naoaki

P-058

Mania

Anastasia

O-006

KurodaTomokoP-083

Marandi

Elahe

P-013

KurodaYasushiO-014

Maruyama

Osamu

O-034

Kuroda

Yuka

O-011

Matsuo

Ryoko

P-084

KusudaTomoyoP-072

Matsumoto

YukikoO-018

Kuwahara

Akira

P-018 P-067

Matsunaga

ShigetakaO-020

Kuwahata

Akiko

P-080

Matsuura

Koji

O-007 O-011

Kuwayama

Masashige

P-033 P-084

Matsuura

ToshikiP-019

Mazloomi

Pooya

O-035

Meddeb

Sawsen

P-070

Mehri

Souhir

P-064 P-070

Memmi

Nedia

P-078

Merdassi

Ghaya

P-078

Metodieva

Meglena. M

P-021

Milyutina

Maria

O-019

Minami

Naojiro

P-002

Mio

YasuyukiO-013 O-015

Mitrani

EduardoO-025

MittalSuneeta

O-038 P-061

Miyazaki

Kohji

P-057

Mizumoto

KumikoP-026

MizutaShimpeiO-014

Mladova

Elena S.

O-019

Moein

Mohammadreza

O-003

Moon

Shin Yong

P-079

Moradkhani

Mojgan

P-053

Mori

Chiemi

P-069 P-033

Morimoto

Yoshiharu

O-024

Morioka

Hitoshi

P-026

Moshiduki

YoshikoO-036

Motoyama

Hiroaki

P-047

Motoyama

Hiroshi

P-041

P-044 O-029

MovagharBahar

P-042 P-049

O-031 P-017

Movahedin

Mansoureh

P-010 P-053

Mowla

Javad

P-059

L
Laufer

Neri

O-025

Lavender

Ben

O-006

LaveryStuartO-006

Lebovich

Meital

O-025

Lee

J.H.

P-025 P-029

Lee

Jaeseok

P-060

Lee

K.H.

P-025 P-029

Levin

Ishai

P-063

Li

Shangwei

P-027

Li

Xiaohong

P-027

Li

Yanping

O-028

Li

Yonggang

O-028 O-029
O-031 P-017
P-044

Li

YunxiuO-028

Liebermann

Juergen

P-068

Lin

Shin-Yu

P-082

Liu

Jiaen

O-005

Liu

Xiaohong

O-005

Luo

Xiu

O-030

Lyv

QiFeng

O-042

M
Ma

Yan ping

Mahdi

Shams-Ara

P-059

Makoolati

ZohrehP-010

312

●Author Index – Oral Communications and Poster Presentations
N

Oki

Chie

O-040 O-041

NAGAI

Yasushi

O-017

Oki

Toshimichi

O-040 O-041

Nagase

Yuki

P-019

OkimuraTadashi

P-069

Nagashima

Hiroshi

P-085

Okubo

Tsuyoshi

P-084

Nagayama

Shiho

P-043

Okutsu

Yuki

P-081

NagoshiKazusuke

P-034

Oride

Aki

P-057

Nakabayashi

Masao

O-033

Osada

Hisao

P-085 P-086

Nakajou

YukikoO-041

Nakaoka

Yoshiharu

O-024

OTSUKI

Junko

O-017

Nakayama

Setsuko

O-033

Ozel

Murat

P-045

Nanbakhsh

FaribaO-035

Narahara

HisashiP-038

NaruseKeiji

O-007 O-011

Park

Chan

P-073

Natalia

Devi

P-074

Park

I.H.

P-029

Nayernia

Karim

P-059

PARK

ILHAE

P-025

Nejatbakhsh

R

O-032

Park

Kyung Eui

P-079

Nihei

Naoki

P-065

Pei

Li

O-030

NikzadHossein

P-003

Penkova

Kalinka. L

P-021

Nishihara

Tomijirou

P-019

PermadiWiryawan

P-074

Nishiyama

Rika

O-027

Pool

Thomas

P-068

NitzschkeMarkusO-037

Prechanich

Japarath

P-036 P-037

Noda

Takahiro

P-005 P-062

Lang

P-027

P-087

P

Q
Qing

O
Obata

Seiichiro

P-028

R

Ochi

Masanori

P-080

Rajaei

FarzadP-001

Ochiai

Keiko

P-085

Ranaei

Ehsan

P-059

Ogata

Seiji

O-018

Ray

PierreP-078

Oh

Sun KyungP-079

Reichart

Aniko

O-021

Ohara

Ken

O-020

Revel

Ariel

O-009 O-025

Ohara

Motohiro

P-041

Rezapour

Sadegh

P-013

Ohhashi

HiroeiP-054

Rezazade V

MojtabaP-032

Ohmomo

Yukio

P-054

Ribic Pucelj

Martina

O-039

Ohori

TadashiP-008

Ripperger

Juergen

P-076

Okada

Hiroshi

O-034

Roh

Sung il

P-060 P-073

Okada

Maki

P-026

Rohani

Mahdi

P-004

Okamoto

Naoki

P-077

Rusa

Ion M

P-056

Okano

Shinichiro

P-072
S

313

●Author Index – Oral Communications and Poster Presentations
Saad

Ali

P-064 P-070

Simon

Alex

O-025

Sahraei

Saeideh

P-049

Singh

Neeta

O-038 P-061

Saito

Hidekazu

P-014

Sipos

Miklos

O-021

Saito

Masahiro

O-020

SoebijantoSoegiharto

O-009

Saito

Takakazu

P-014

Song

Sang Jin

P-016

Sakaguchi

Yuuko

P-026

Srivastava

Maya D

P-012

SAKAI

K

O-008

Stefanescu

Coralia V.

P-056

Sakamoto

Hideki

O-033

Stevens

JulietO-009

Sakhinia

Ebrahim

P-004

Su

Yi-Ning

P-082

SakuraiTomoyoshi

O-004

Suenaga

Manami

O-022

Salehnia

Mojdeh

P-013

SueokaKou

O-004 P-058

Salem

RifaatO-005

Sugimura

Satoshi

P-071

Salem

Shala

O-005

Sugishita

Yodo

P-077

SANKAI

Tadashi

O-017

Sun

H.G.

P-025 P-029

Sato

Kenji

O-004

Supriyadi

Agus

P-040

Sato

SetsukoO-012

Susanti

Marly

O-010

Sato

SuguruO-004

Suzuki

Masakuni

P-005 P-023

Sato

YasukoO-022

P-043 P-046

Sawada

Tomio

P-051

P-062 P-066

SegawaTomoyaP-084 P-087

Suzuki

Tatsuya

P-023 P-043

Seki

Hiroyuki

O-020

Swain

Jason E

P-068

Seol

Hye WonP-079

ShahaliM

P-039 P-059

Shahhoseini

MaryamP-049

Tabibnejad

Nasim

O-003

ShahirMohammadP-003

TachibanaIkuo

P-005 P-046

Taherian

AliakbarP-003

Takahashi

Kaori

O-004

Takahashi

NoriyukiP-077 P-081

Takahashi

Sigetomo

P-005

Takai

YasushiO-020

Takamura

Akina

P-072

TAKANO

Jun-ichiro

O-017

TakasakiAkihisa

P-026

Takayama

Yuko

P-033

TakedaYoshiharu

O-033

Takehara

Yuji

P-033 P-035

T

Hossein
Sharbatoghli

Mina

P-032

Sheng

Jian-Zhong

O-016

Shibahara

Hiroaki

P-023 P-043

SHIBUYAH

O-008

ShimadaKazuhiko

P-043

Shimada

Naoko

P-084

Shimamura

Katsunori

P-026

Shimizu

Miwa

P-014

Shimizu

Yasufumi

P-041

Shimoi

Gaku

P-011

Shiotani

MasahideO-014 O-018

Shoda

Akiko

O-036

P-069 P-083
P-085 P-086

314

●Author Index – Oral Communications and Poster Presentations
P-087

WagmanIsrael

P-063

Takeuchi

Masayoshi

O-026

Wang

Hong-Hua

P-007

Talebi

Saeed

P-055

Wang

Yun

O-042

TanakaKohei

P-062 P-066

Watanabe

Aiko

O-026 O-027

P-046

WEISS

B

P-052

TanakaYu

P-018 P-067

Wells

Dagan

O-006

Tang

Li

O-016

Wiweko

Budi

O-009

Tang

Xiaohui

O-033

Wu

Yan-Ting

O-016

Tani

Hirohiko

P-014

Wu

Ze

O-028 O-029

Tarumi

WataruP-081

O-031 P-044

Tatsumi

Ken-Ichi

O-034

P-017

Terai

Kazutaka

O-034

Teramoto

Shoukichi

P-084

Tirtajasa

Caroline

O-009

Tochigi

HidenoP-036 P-037

Y

Torfeh

Mahnaz

P-004

Trew

Geoffrey

Tsukamoto

X
Xiang

Jing-Ying

P-007

YabuuchiAkiko

P-033 P-085

O-006

YamadaMasayasu

P-002

Sanae

P-081

YamadaSatoshiO-018

TsunodaSatoshiO-022

YamadaYoshiakiP-008

Tulandi

Togas

P-063

Yamadera

Rie

P-083

Yamagata

Kazuo

O-024

Yamamoto

Yuri

P-018 P-067

Yamanouchi

Tadayuki

P-071

Yamasaki

Hideki

O-040

Yang

Xinzhi

P-072

Yang

Zhihong

O-005

Yano

Yuya

P-018 P-067

YASMIN

Lubna

O-017

Ye

Hong

O-030

Yekta

Zahra

O-035

U
Uchiyama

Kazuo

P-035

Ueno

Satoshi

P-035

Uhereczky

Gabriella

O-021

Urano

Akiyoshi

P-014

Usui

Mitsuru

P-019

UtsumiTakanobu

P-065

Utsunomiya

Yufuko

P-038

V
Vajta

Gabor

O-021

Yoav

Smith

O-009

Virant Klun

Irma

O-039

YokotaHidemi

O-012

VIVIERC

P-052

YokotaMikakoO-012

VoglerAndrej

O-039

YokotaYoshimasa

O-012

Yorimitsu

Takeshi

P-041

Yoshikai

Kaori

P-051

Yoshimura

Yasunori

P-058 O-004

Yoshioka

Maiko

P-019

W
WACHI

Y

O-008

Wada

Tomohisa

P-080

315

●Author Index – Oral Communications and Poster Presentations
Yoshioka

Nobuhito

P-077

Youssef

KhaledP-064

Yu

Hong-Ying

P-007

YumotoKeitaro

O-013 O-015

Yun

Ji-HyunP-015 P-016

ZarnaniAmir Hassan

P-055

Zeng

Pinghong

O-030

Zeng

Xun

P-027

Zhang

John

O-042

Zhang

Xiaomei

P-024

Zhioua

Amel

P-078

Zhioua

Fethi

P-078

Zhou

Tian-Hua

O-016

Zhou

Xiao-Jin

P-007

Z

316

●Certificate of AttendanceInternational Society for
In Vitro Fertilization

International Society for In Vitro Fertilization

Certificate of AttendanceThis document certifies that

attended the16th World Congress on In Vitro Fertilization &6th World Congress on In Vitro Maturation
September 10-13, 2011, Tokyo, Japan
Signature of Participant

René Frydman
Society President

317

Osamu Kato
Congress President