Introducing reagents for use in
rat reproductive engineering- A medium for rat in vitro fertilization -Massive improvements in rat strain preservation
efficiency and live animal maintenance costs
through rat sperm cryopreservation!

Manufacturer

Distributor

Technical Support :
fertiup@kyudo.co.jp

E-mail : info@cosmobiousa.com
URL : https://www.cosmobiousa.com

・ Are you having trouble with the preservation ofgenetically modified rat strains?
・ Are you worried about the cost of maintaining
and managing these rat strains?
Then rat sperm cryopreservation techniques could bethe answer to your problems!
Try CARD reagents for rat reproductive technology.
Rat sperm cryopreservation techniques - Development background
Rats are widely used in medical and life science research. In recent years, various strains of genetically modified rat havebeen produced worldwide thanks to the development of genome editing technologies. Moreover, it is predicted that evenmore genetically modified rats will be produced in the future. Therefore, there is a pressing need for effective preservation
techniques for these genetically modified rat strains.
Through sperm freezing, it is possible to quickly cryopreserve a large number of sperm taken from the cauda epididymis
of a male rat. For this reason, sperm cryopreservation is expected to be an easier, more effective and most economically
viable technique for the preservation of genetically modified rat strains than embryo cryopreservation. However, rat
sperm is vulnerable to the effects of physical changes to pH, osmotic pressure, temperature and other similar factors.
Moreover, it is difficult to maintain the motility of frozen-thawed sperm via conventional sperm cryopreservation methods,
which means that it is not viable to use these sperm in in vitro fertilisation (IVF). Therefore, the establishment of rat
sperm cryopreservation and rat IVF techniques was required to solve these issues.
A research team led by Dr. Naomi Nakagata and Dr. Toru Takeo at the Institute of Resource Development and Analysis,
Kumamoto University have developed a practical rat sperm cryopreservation technique and an IVF method using frozenthawed rat sperm. These techniques may offer a solution to the problem of genetically modified rat strain preservation.

Reagents and equipment for use in rat reproductive engineering
CARD FERTIUP® Rat Sperm Cryopreservation AgentCARD FERTIUP🄬 Rat Sperm Cryopreservation Agent is a newly-developed agent which enables researchers toefficiently carry out cryopreservation of rat sperm, which until now has been notoriously difficult to cryopreserve.
Conventional rat sperm cryopreservation methods are unable to effectively maintain the motility of frozen-thawed sperm.
On the other hand, rat sperm cryopreservation carried out using CARD FERTIUP🄬 Rat Sperm Cryopreservation Agentyields superior motility even in frozen-thawed rat sperm, allowing for the production of many rat pups via IVF andembryo transfer.

CARD Rat IVF medium
CARD Rat IVF Medium is a new IVF medium developed specially for use in rat IVF.

CARD Rat sperm freezing kit
CARD Rat Sperm Freezing Kit is a kit that facilitates the effective production of high-quality frozen rat sperm during rat
sperm cryopreservation using CARD FERTIUP🄬 Rat Sperm Cryopreservation Agent. With this kit, researchers canappropriately carry out the rat sperm cooling process during rat sperm cryopreservation.

Characteristics and effectivenessRat sperm cryopreservation using CARD FERTIUP🄬 Rat Sperm Cryopreservation Agent and rat IVF carried out using
frozen-thawed sperm are revolutionary techniques which have enabled the production of high quality frozen rat sperm
and the acquisition of high IVF efficiency rates using frozen-thawed rat sperm at a practical level for the very first time.
The fertilised oocytes obtained via IVF develop well, and after transferring embryos to a female rat, one can efficiently
acquire genetically modified rat pups. (see Table 1, Table 2 and Fig. 1)

Usage examples / benefits
1． Implementation of high quality IVF using frozen-thawed rat sperm
2． Storage of genetically modified rat strains for subsequent use in the production of pups
3． Massive reduction in rat maintenance costs through effective rat strain preservation

12Fresh
34Total
12Frozen 3
4Total

198 s
183 s
195 s
182 s
758 s
161 s
191 s
201 s
196 s
749 s

No. offertilised
oocytes

Fertilisation
rate (%)

185 s
170 s
181 s
174 s
710 s
135 s
160 s
161 s
159 s
615 s

93.4 s92.9 s92.8 s
95.6 s
93.7±1.3 s
83.9 s83.8 s80.1 s81.1 s
82.1±1.9 s

93.7%±1.3

100Fertilization rate (%)

Sperm No.

No. ofinseminatedoocytes

82.1%±1.9
80
60
40200

Fresh sperm

Table1 / Fig.1

IVF fertilisation rates using fresh and frozen-thawed rat sperm.

Sperm

No. of 2-cellembryos

No.

12
Fresh
3
4
Total
12
Frozen34
Total

No. ofblastocysts

100 s
100 s
100 s
100 s
400 s
100 s
100 s
100 s
100 s
400 s

58 s
63 s
65 s
71 s
257 s
72 s
57 s
64 s
53 s
246 s

Developmentalrate(%)

Fertilised oocytes
transferred

58 s
63 s
65 s
71 s
64.3±5.4 s
72 s
57 s
64 s
53 s61.5±5.4 s

20 s
20 s
20 s
20 s
80 s
20 s
20 s
20 s
20 s
80 s

Frozen/Thawed sperm

No. of pups
13 s
12 s
13 s
10 s
48 s
12 s
12 s
10 s
12 s
46 s

Birthrate(%)
65.0 s60.0 s65.0 s50.0 s60±7.1 s
60.0 s60.0 s50.0 s60.0 s
57.5±5.0 s

Table2 Rate of development of fertilised oocytes derived from fresh and frozen-thawed rat sperm to the blastocyst stage,
and birth rate of said fertilised oocyte after oocyte transfer
Reference : Naomi Nakagata, Nobuyuki Mikoda, Satohiro Nakao, Ena Nakatsukasa & Toru Takeo, Establishment of sperm
cryopreservation and in vitro fertilisation protocols for rats. SCIENTIFIC REPORTS, 10:93, 2020.

Rat sperm

Rat unfertilized Rat pronuclear
oocyteoocyte

Rat 2-cellembryos

Rat blastocysts

Live pups derived from in vitro fertilisation
with cryopreserved rat sperm

The above data and photographs are published with the permission of Kumamoto University.

Reproductive Engineering Reagent List
Code No.

Reagents for use in rat reproductive engineering
🄬KYD-FR-001

NEW CARD FERTIUP

KYD-FR-002

NEW CARD Rat IVF medium- A medium for rat in vitro fertilization -KYD-GAR-001

NEW CARD Rat sperm freezing kit

Code No.
KYD-001-05-EX
KYD-001-05-EX-X5
KYD-001-EX
KYD-001-EX-X5
KYD-002-05-EX
KYD-002-05-EX-X5
KYD-002-EX
KYD-002-EX-X5Rat Sperm Cryopreservation

Reagents for use in mouse reproductive engineering
CARD FERTIUP® Mouse Sperm Cryoroprotectant : CPA

CARD FERTIUP® Mouse Sperm Preincubation Medium : PM

STANDARD
1×1.5 mL
1×1.0 mL
1 kit

STANDARD
1×0.5 mL
5×0.5 mL
1×1.0 mL
5×1.0 mL
1×0.5 mL
5×0.5 mL
1×1.0 mL
1×1.0 mL

KYD-003-EX

CARD MEDIUM® Mouse Fertilization Medium

1 kit

KYD-004-EX

CARD FERTIUP® PM (1.0 mL) and CARD MEDIUM®

1 set

KYD-005-EX

CARD FERTIUP® PM (0.5 mL) and CARD MEDIUM®

1 set

KYD-010-06-EX
KYD-010-06-EX-X5
KYD-010-EX
KYD-010-EX-X5
KYD-015-EX
KYD-015-EX-X5
KYD-008-02-EX
KYD-008-02-EX-X5
KYD-008-05-EX
KYD-008-05-EX-X3
KYD-011-EX-2
KYD-011-EX-5KYD-012-EX
KYD-013-EX-0.5
KYD-013-EX-1
KYD-014-EX-2
KYD-014-EX-5

CARD HyperOva® Superovulation Reagent for Mice

CARD HyperOva® F.D. Superovulation Reagent for Mice

CARD mHTF

CARD KSOMCARD 1M DMSOCARD DAP213
CARD 0.25M Sucrose

1×0.6 mL
5×0.6 mL
1×1.0 mL
5×1.0 mL
1×2.0 mL
5×2.0 mL
1×2.0 mL
5×2.0 mL
1×5.0 mL
3×5.0 mL
1×2.0 mL
1×5.0 mL
1×1.0 mL
1×0.5 mL
1×1.0 mL
1×2.0 mL
1×5.0 mL

Code No.

Accessory for use in mouse/rat reproductive engineering STANDARD

KYD-006-EX

CARD Cold Transport Kit

1 Set

KYD-S036

Embryo manipulation instrument set

1 Set

KYD-S018

Freezing Canister

Unit

KYD-S035

Triangular Cassette Long (10units)

10 Units

KYD-S020X10

Sperm Straws (10 Pieces x 10 Units)

10 Units

Note : CARD reagents for mouse/rat reproductive technology are for laboratory use only and are not to be
used for the diagnosis or treatment of illnesses.
※FERTIUP is our registered trademark.