KYUDO“best reagents” for “best research”

IVF

-19６℃

Cryopreservation

Embryo Transfer

Cutting-edge technology brought to youmouse
Cold Transport Kit♂

Mouse Sperm
Cryoprotectant
Agentmouse♀

CARD HyperOva F.D.
CARD HyperOva

P7

P3
P24℃

Mouse Sperm-196℃

Cold storage

Cryopreservation

Preincubation Medium

Superovulation
1M DMSO

P2

Sperm preincubation

DAP213

CARD MEDIUM
ｍHTF

P4
-196℃

P2・P4

1M DMSO

Cryopreservation

In vitro fertilization

0.25M

DAP213

SucroseP4P4KSOM-196℃

Cryopreservation

Embryo culture
0.25M

SucroseP4P４Embryo transfer

P7Birth of pups

P1

Reagents for Reproductive Technology of Mouse
CARD FERTIUP Mouse Sperm Cryoprotectant AgentCARD FERTIUP Mouse Sperm Cryoprotectant Agent mitigates freezing damage
during sperm cryopreservation and improves in vitro fertilization rates using
frozen-thawed sperm in numerous mouse strains, including the C57BL/6 strain.
This agent also facilitates a massive reduction in costs associated with theupkeep of strains in comparison to the cost of live animal maintenance.(①)＊Reference(p９)：No.1

CARD FERTIUP Mouse Sperm Preincubation Medium
CARD FERTIUP Mouse Sperm Preincubation Medium promotes
capacitation of mouse sperm and improves in vitro fertilization
rates using fresh, frozen-thawed and cold stored mouse sperm.(②)＊Reference(p９)：No.2＊Patent(JP:4683408）

CARD MEDIUM Mouse Fertilization Medium
CARD MEDIUM High-Performance Mouse IVF Medium brings about amassive improvement in the in vitro fertilization rates of numerous mousestrains, including the C57LB/6 and 129 strains. In fact, using CARD
MEDIUM alongside CARD FERTIUP Mouse Sperm Preincubation Medium
can yield in vitro fertilization rates of over 80% for frozen-thawedC57BL/6 mouse sperm.(③)＊Reference(p９)：No.3＊Patent（JP:5439677|US:9,994,868 B2）
Fertilization rate of frozen-thawed C57BL/6 sperm

obtain fertilization rates of over 80%

using frozen-thawed C57BL/6 sperm!

Fertilization rate（％）

With CARD MEDIUM, we can even

89.2%

100（①）
（②）
（③）

８０
６０
４０
２０

17.9%

０R18S3+HTF

CARD FERTIUP+CARD MEDIUM

P2

CARD HyperOva Superovulation Reagent for Mouse
CARD HyperOva Mouse Superovulation Reagent enables us toobtain a much larger number of oocytes from females of numerous
mouse strains than via conventional superovulation methods.
CARD HyperOva may bring about a threefold or even fourfold
reduction in the required number of donor mice. This contributes
to a reduction in the number of animals per experiment, thereby
upholding the 3 Rs Principle(④)＊Reference(p９)：No.4・No.5＊Patent（JP:5927588｜US:10,500,275,B2｜EU:3260535）

CARD HyperOvaF.D. Superovulation Reagent for Mouse
CARD HyperOva F.D. retains the high performance ofCARD HyperOva while also oﬀering signiﬁcantly improved
storage stability through freeze drying. After preparing thereagent, cryopreserve it at a temperature at -20ºC orbelow, and then take as much of the reagent as you need,whenever you need it.(⑤)

Using CARD HyperOva, we can obtain three to four times more

oocytes than conventional superovulation methods in C57BL/6 mice!

107

140

Number ofovulated oocytes

120
100

４００

PMSG+hCG■

CARD HyperOve+hCG

68

52

８０６０

■

90

2724

29

28

7２０P3（④）（⑤）

C57BL6/J

A/J

BALB/cByJ

C3H/HeJ

22DBA/2J

1625

FVB/NJ2533

CD1CARD mHTF medium
CARD mHTF Medium is an in vitro fertilization
medium. This medium enables superior in vitro
fertilization rates and a high yield of 2-cellembryos.

CARD KSOMCARD KSOM is a culture medium
for use on embryos obtained via

in vitro fertilization. CARD KSOM
contributes to superior development
rates from 2-cell embryos to blastocysts.

CARD １M DMSOCARD 1M DMSO is cryopreservation
reagent utilized in simple vitriﬁcation.
It is used in the cryopreservation ofembryos and oocytes.

CARD DAP213

CARD DAP213

CARD DAP213 is cryopreservation
reagent utilized in simple vitriﬁcation.
It is used in the cryopreservation ofembryos and oocytes.

CARD 0.25M Sucrose
CARD 0.25M Sucrose is used to warm
vitriﬁed embryos and oocytes via simple
vitriﬁcation.

P4

Breaking new boundaries - Our new technology
Rats

♂

Rats

♀

CARD FERTIUP Rat Sperm
Cryopreservation Agent

P6-196℃

Cryopreservation
Rat Sperm Freezing Kit

P7

Rat IVF Medium

Superovulation
P6

In vitro fertilization

Embryo culture

Embryo transfer
P7
Birth of pups

P5

Reagents for Reproductive Technology of Rats
CARD FERTIUP Rat Sperm Cryoprotectant AgentCARD FERTIUP Rat Sperm Cryopreservation Agent is the worldʼs ﬁrst practicalcryopreservation agent for use with rat sperm. Previously rat sperm cryopreservation
was an extremely diﬃcult task, but CARD FERTIUP Rat Cryopreservation Agent can
now make it possible to cryopreserve rat sperm in an eﬃcient manner. Whatʼs more,
once thawed the rat sperm maintain a high motility and can be used in in vitro
fertilization.

◆Store at -20℃ or below.
◆For laboratory use only. Not to be used for diagnosis or treatment of illnesses.
◆This product improves the efficacy of rat sperm cryopreservation.
After thawing, rat sperm can be used for in vitro fertilization.

＊Reference(p９)：No.6【Manufacturer】

For use on rats only.

CARD Rat IVF Medium
CARD Rat IVF Medium is a novel medium
developed speciﬁcally for use in rat in vitro
fertilization. It oﬀers superior in vitro
ferdtilization rates, particularly when using
frozen-thawed rat sperm.

We can obtain high fertilization rates using
frozen-thawed sperm in several rats strains!

Frozen-thawed rat sperm

Fertilization rate（％）

10095.8%＊Reference(p９)：No.7

93.5% 90.7%

94.5%

60.2%

８０６０

25.7%

４０２００CD (SD)

BNLE

F344

Wistar

EGFP

P6

Reproductive Technology Tools for Mouse/Rat（A）

CARD Cold Transport Kit

（C）

（D）

The CARD Cold Transport Kit provides everything
you need to safely and cheaply transport mousecauda epididymides and 2-cell embryos at refrigerated
temperatures. The transported cauda epididymides
and embryos can used at the destination institution to
produce live mice via reproductive engineering
techniques.

（B）
Usage:
Transport of mouse cauda epididymides and2-cell embryos at refrigerated temperatures.
Cold storage retention period:
Max. 72 hours(when using our cold storage solution for cauda epididymides)
Kit contents:
A cold storage box (A), cold packs (B; 4 x large, 2 x small),a Thermos bottle (C), paper boxes and packing material (D).

CARD Rat Sperm Freezing Kit
CARD Rat Sperm Freezing Kit is a useful kit for producing
high-quality frozen rat sperm using CARD FERTIUP Rat
Sperm Cryopreservation Agent. It allows users to correctly
carry out the cooling process during the sperm（A）

（B）
（C）

cryopreservation operation.

Kit contents:
A Styrofoam box (A), sperm cryopreservation straw
holding support (foam ﬂoat (B) and plate (C))

Embryo Manipulation Instrument Kit（D）

Our embryo manipulation instrument kit utilizes a glass capillary toensure the uniform transfer of embryos.

（C）

（B）

（E）

Set contents: Glass capillaries for embryo transfer (20 pieces; dry-heatsterilized) (A), a connector (B), a Pasteur pipette(C), a mouthpiece (D),a silicon tube (E)* We will send customers all equipment required for embryo transfer.

P7（A）

Straw Connector
A straw connector is used for sucking
up sperm suspended in CARD FERTIUP
Sperm Cryopreservation Agent into
sperm straws.

Triangular Cassette Long (W278mm)
Triangular Cassette Short (W145mm)
Cassettes used when cryopreserving straws containing sperm
suspension in liquid nitrogen. We oﬀer two types of cassette: along 278mm cassette, which can store up to around 30 straws,
and a short 145mm, which can store up to around 15 straws.

Sperm Freezing Straws (10 pieces)This is a 0.25mL straw used for sperm storage.
Our straws enable the storage of sperm inliquid nitrogen for long periods of time. Allstraws have been sterilized using EOG.

Freezing Canister
A freezing canister for use in sperm
cryopreservation. Using a ﬂoat guarantees
a cooling rate conducive to mouse sperm
cryopreservation on liquid nitrogen, which
in turn enables the storage of higher
quality frozen sperm.

P8

About Purchases from Outside of Japan
Our reproductive technology reagent products areused by research institutions around the world.

2792 Loker Avenue West, Suite 101
Carlsbad, CA 92010 USA
https://www.cosmobiousa.com/
info@cosmobiousa.com

HIBIYA CENTRAL BLDG, 1-2-9 Nishi
Shimbashi, Minato-ku, Tokyo
105-0003, Japan
http://www.nomjim.co.jp/en/contact/

11-6 Hongo 6-chome, Bunkyo-ku,Tokyo 113-0033, Japan
http://www.fujihira.co.jp/english/contact.html

Technical support
If you have any questions, queries or issues concerning our reagents for use in mouse/rat
reproductive technology, please feel free to contact us using the contact details below.
Our veteran technicians are on hand to help solve any problems that you may have.

web_req@kyudo.co.jp

References
Reference

No.
1

P9

Takeo T, Nakagata N. Combination medium of cryoprotective agents containing L-glutamine and methyl-β-cyclodextrin in a preincubation medium yields a high fertilization rate for cryopreserved C57BL/6J mouse sperm. Laboratory Animals, 44(2), 132-137, 2010

2

Takeo T, Hoshii T, Kondo Y, Toyodome H, Arima H, Yamamura K, Irie T, Nakagata N. Methyl-beta-cyclodextrin improves fertilizing
ability of C57BL/6 mouse sperm after freezing and thawing by facilitating cholesterol eﬄux from the cells. Biology of Reproduction, 78(3):546-51, 2008

3

Takeo T, Nakagata N. Reduced glutathione enhances fertility of frozen/thawed C57BL/6 mouse sperm after exposure to methyl-beta-cyclodextrin. Biology of Reproduction, 85(5), 1066-1072, 2011

4

Takeo T, Nakagata N. Immunotherapy using inhibin antiserum enhanced the eﬃcacy of equine chorionic gonadotropin on super-5

Takeo T, Nakagata N. Superovulation using the combined administration of inhibin antiserum and equine chorionic gonadotropin

6Nakagata N, Mikoda N, Nakao S, Nakatsukasa E, Takeo T. Establishment of sperm cryopreservation and in vitro fertilisation pro-7

Takeo T, Nakao S, Mikoda N, Yamaga K, Maeda R, Tsuchiyama S, Nakatsukasa E, Nakagata N. Optimized protocols for sperm

ovulation in major inbred and outbred mice strains. Theriogenology, 86(5), 1341-1346, 2016

increases the number of ovulated oocytes in C57BL/6 female mice. PLoS One, 10(5), 2015

tocols for rats. Scientiﬁc Reports, 10, 93, 2020

cryopreservation and in vitro fertilization in the rat. Lab animal, 51, 256-274, 2022

Products List
STANDARD

Storage Temp.

CARD FERTIUP Rat Sperm Cryoprotectant Agent

1.5 mL-80℃ to -20℃

CARD Rat IVF Medium

1.0 mL４℃

STANDARDStorage Temp.

Reagents for Rat Reproductive Technology

Reagents for Mouse Reproductive Technology
CARD FERTIUP Mouse Sperm Cryoprotectant Agent: CPA

CARD FERTIUP Mouse Sperm Preincubation Medium: PMCARD MEDIUM Mouse Fertilization Medium
CARD FERTIUP PM (0.5mL) and
CARD MEDIUM Mouse Fertilization Medium Kit
CARD FERTIUP PM (1.0mL) and
CARD MEDIUM Mouse Fertilization Medium Kit
CARD HyperOva Superovulation Reagent for Mouse

CARD HyperOva F.D. Superovulation Reagent for Mouse
CARD ｍHTF Medium

CARD KSOMCARD 0.25M Sucrose
CARD 1M DMSOCARD DAP213

0.5 mL
1.0 mL
0.5 mL
1.0 mL
1 kit

18 to 28℃

4℃
4℃

1 set
4℃
1 set
0.6 mL
1.0 mL
1 kit
5 kit
2.0 mL
5.0 mL
2.0 mL
5.0 mL
2.0 mL
5.0 mL
1.0 mL
0.5 mL
1.0 mL-80℃ to -20℃

4℃

4℃

4℃

4℃
4℃
4℃

STANDARD

Storage Temp.

Straw Connector

1 unit-Triangular Cassette Short (W145mm)

10 units-Triangular Cassette Long (W278mm)

10 units-

Sperm Freezing Straws (10 pieces)

10 units-

1 kit

4℃*Embryo Manipulation Instrument Kit

1 kit-Freezing Canister

1 unit-1 kit-Reproductive Technology Tools for Mouse/Rat

CARD Cold Transport Kit(*The storage temp. of cold storage solution for cauda epididymis)

CARD Rat Sperm Freezing Kit

Mouse/Rat Reproductive Engineering Reagents are intended for research use only and are not intended for diagnostic or therapeutic purposes.
FERTIUP, CARD MEDIUM, and CARD HyperOva are registered trademarks.
All information is current as of October 2022.

P10

Are you having trouble
storing your transgenic
mouse or rat strains?
Are you struggling withmaintenance, upkeep orbreeding?
We can help.
We at Kyudo Co, Ltd. provide a wide range of reagents foruse at every stage of mouse and rat reproductive technology,
from superovulation and in vitro fertilization through to thefreezing of sperm and embryos.

These high quality reagents were developed in collaboration
with the Center for Animal Resources and Development,
Kumamoto University. We manufacture and sell the reagents
under license to customers worldwide.

Using reproductive engineering techniques alongside ourreagents contributes to a signiﬁcant reduction in the numberof animals required per experiment, thereby following
the 3R Principle.

Kyudo Co., Ltd