Quality Results for life

PRODUCT CATALOGUE

Oligospermia
Cryopreservation〇 Freeze and thaw kit for low-count sperm and oligospermia
using Cryotop®.
〇 Sucrose is the cryoprotectant used in the sperm freeze solution,and no Glycerol is used.
〇 Post-thaw sperm motility and viability can also be evaluated with the kit.
〇 Cryotop® -wide is available on its own in packs of 10.
(REF. 81016, 81017, 81018, 81019 and 81020)
Collaborative Development: Atsushi TANAKA, MD., St. Mother Hospital Infertility Clinic / Kitazato Corporation

REFCodeContents92231

Oligospermia Cryo KitCryotop®-wide（white)×2
Collagenase 0.5 mL×1

Sperm Medium 0.5 mL×1
Sperm Freeze（SF4) 0.5 mL×192232

Oligospermia Thawing Media

Sperm Medium 0.5 mL×1
Pentoxifylline 0.5 mL×1

HOST Solution 0.5 mL×1
PVP3％ 0.5 mL×1

COMPONENTSQUALITY CONTROL

Calcium Chloride / Collagenase / Gentamicin / Glucose /pH 7.2-7.6 / Osmolarity / Endotoxin <0.25EU/mL / Sterility Test / Mouse Embryo Assay ≥80%

HEPES / Human serum albumin / Magnesium sulfate /Sperm Freeze（SF4）: Sperm Survival ≥80% / Sperm Cryosurvival ≥0.5Pentoxifylline / Polyvinylpyrrolidone / Potassium chloride /PVP3% : Sperm Motility Index

Potassium phosphate / Sodium bicarbonate / Sodium chloride /Sodium lactate / Sodium pyruvate / Sucrose

Pentoxifylline : Sperm Survival (24h) ≥75% / Sperm Penetration ≥3
Storage：2-8℃

Shelf Life：6 months (Collagenase)

RESULT
Classified
Spermatozoa

No. ofpatient

No. ofET cycle

Sperm ※1
colleted rate

Sperm ※2 2PN fertilization Average number of Average number ofembryos transferredfrozen embryos
survival rate
rate (N)（min-max)（min-max)

Frozen ejaculatedspermatozoa

2860

97.8%
[510/521]87.1%[444/510]a52.7%[224/425]b

1.52
（1-2)

0.72
（0-1)

Frozen TESE
spermatozoa

2018

92.7%
[152/164]60.5%[92/152]a’

37.2%[29/78]b’

1.73
（1-2)

0.53
（0-1)

Fresh ejaculated
spermatozoa

31107

No data

No data

52.2%[302/579]

1.41（1-2)

1.83
（0-4)

a-a’ and b-b’ P<0.05 （Chi-square test)※1 collected spermatozoa/ warmed spermatozoa※2 survived spermatozoa / collected sperm

From St. Mother Hospital Infertility Clinic

REFERENCE〇 Endo et al. Clinical and neonatal outcomes of individually vitrified human sperm with Cryotop and Cell Sleeper. Cryobiology. 2022 Oct:108:78-81.
Specification may change without pre-notice for purpose of product improvement.

Kitazato Corporation
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Mail

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PROTOCOL

Kitazato corporation
Official YouTubeThe video protocol is available
on our official YouTube channel.

Collection of low-count sperm and Freeze procedure
12With Collagenase from the Oligospermia Cyro Kit, make a drop for sperm
collection on a glass bottom dish.
Place the collected seminiferous tubules in the drop and cut theminto small pieces using a 27G needle.
*For ejaculated sperm, start from 2 .

Drop for aspirated sperm
Collagenase (30-50µL)

TESE pick up pipette

Aspirate good sperm with a TESE pick-up pipetteand transfer them into a fresh sperm collection
drop of Sperm Medium.

Drop for sperm

Drop for aspirated sperm
Sperm Medium

3Sperm Medium (Approx. 1µL)

Place both Cryotop® and the dish under an inverted microscope so that both are visible.
Make a 2µL drop of Sperm Freeze on the Cryotop®.
TESE pick up pipette
TESE pick up pipette

Aspirate sperm from its tail with the TESE pick-up pipette andquickly transfer the sperm into the Sperm Freeze drop.
Immediately place the Cryotop® on Sperm Freeze Board
2cm above LN2 and expose it to the vapour for 2min to freeze.
Sink the Cryotop® in LN2 before cryopreservation
Drop for sperm collection
Sperm Medium (Approx. 1µL)
in a LN2 tank.

Sperm Freeze（2µL）

Cryotop®

Inverted microscope lensTechnique of low-count sperm thawing
45Prepare a dish for sperm thaw and ICSI
as the figure in the right shows.
Equilibrate the dish in a 5% CO2 incubator
for at least 2hr.

Mobility check drop
Pentoxifyllline（2µL）

Thaw drop

Survival check drop

ICSI drop

Washing drop

Sperm Medium（2µL）
Sperm Medium（50µL）

HOST Solution（50µL）
PVP3%（50µL）

Take out Cryotop® from LN2 and air-thaw for 2sec.
Immerse the tip of the Cryotop® in the Sperm
Medium drop on the dish.
If sperm motility is confirmed, perform ICSI
immediately.

Thaw drop

Sperm Medium（2µL）

ICSI

Sperm Medium（50µL）〇 If the sperm was immotile, check its motility inthe Pentoxifylline drop. Perform ICSI after its motility
confirmation.

Mobility check drop
Pentoxifyllne（2µL）

〇 If the sperm remained immotile in the Pentoxifylline drop,
check its viability in the HOST Solution drop.
Perform ICSI after its viability is confirmed.

Survival check drop
HOST Solution（2µL）

HOST（50µl）
HTF with HAS