Assisted Reproductive Technologies

CONTENT:

1. Introduction
2. Overview
3. CARD MEDIUM® und FERTIUP®
4. CARD HyperOva® the superovulation reagent
5. Peripheral products
6. Products

Introduction

In recent years, the number of genetically engineered mice being produced has increased dramatically. Moreover, the rapid progress in the development of genome-editing (TALEN and CRISPR/Cas9) techniques for molecular biology reserach has been remarkable, so much so that a genetically engineered mouse strain can be produced easily in a few months. Production has been supported by reproductive engineering techniques such as In vitro fertilisation, embryo and sperm cryopreservation and embryo transfer techniques. Such techniques have become invaluable peripheral technologies and their use is expanding rapidly. 

Overview

To support your research we offer several reagents and disposable items (general labware) for In vitro fertilisation, In vitro culture, cryopreservation and In vitro manipulation.

CARD MEDIUM® and FERTIUP®

Sperm cryopreservation is a practical and cost-effective method of archiving mouse strains in the laboratory. FERTIUP® and CARD MEDIUM® are high-performance media for mouse sperm cryopreservation, sperm preincubation and In vitro fertilization (IVF). 

When used in combination, FERTIUP® and CARD MEDIUM® provide a high and stable fertilization rate using frozen-thawed mouse sperm. This will allow you to reproduce mice whenever you need them.
The FERTIUP® sperm cryoprotectant and sperm preincubation medium greatly improve fertilization rates of frozen-thawed sperm in C57BL/6 and 129 Mice strains, from less than 10% (using conventional methods) to more than 30%. 
In addition, CARD MEDIUM® enhances the fertility rate to over 80%. FERTIUP® and CARD MEDIUM® have no reproduction toxicity of any kind. 

The newly developed system for sperm cryopreservation and IVF using FERTIUP® and CARD MEDIUM® provides superior support for the safe and reliable preservation of your valuable mouse strains.

No.	Strains	Method	Number of oocytet donors	Number of inseminated oocytes	Average number of ovulated oocytes/female	No. of fertilized eggs	Fertilization rate (%)
1	C57BL/6J	eCG Product	10 10	270 1072	27.7±5.4 107.2±22.7	267 963	96.4±3.2 89.8±3.7
2	BALB/c ByJ	eCG Product	10 10	293 903	29.3±8.9 90.3±14.5	213 680	73.4±11.3 76.4±9.0
3	C3H/HeJ	eCG Product	10 10	286 520	28.6±8.4 52.0±18.1	251 446	87.8±22.6 85.8±15.5
4	DBA/2J	eCG Product	10 10	225 688	22.5±7.8 68.8±13.6	194 592	86.2±11.0 86.0±4.5
5	FVB/NJ	eCG Product	10 10	168 256	16.8±3.3 25.6±5.9	159 239	94.6±6.7 93.3±6.6
6	CD1	eCG Product	10 10	202 337	20.2±8.0 33.7±9.9	146 269	72.3±16.0 79.8±19.6

Fig. 2:

• 7.5 IU of CARD HyperOva® or PMSG was injected to four weeks old female mice, then 7.5 IU of hCG was administered 48 hours later.
  Thereafter all ova obtained 17 hours after the hCG injection were collected.
• The In vitro fertilization rate is defined as the number of 2-cell embryos divided by the total number of ova (mean±SD).
• Data cited from a paper published by Prof. Naomi Nakagata and Toru Takeo (Center for Animal Resources and development, Kumamoto University)

Fig. 3: Fertilization rates using FERTIUP® Preincubation Medium and CARD MEDIUM® for fresh and frozen sperm in various strains.

Fig. 4: Fertilization rates using frozen-thawed sperm with FERTIUP® and with FERTIUP® and CARD MEDIUM®

CARD HyperOva® the superovulation reagent

Reduction in experimental animal use (Contribution to 3Rs):
Using CARD HyperOva® enables us to reduce the number of female mice used for ovum collection by 25 - 30 %.

Optimization of genetically modified mouse creation and preservation:
More ova can be obtained from less female mice using CARD HyperOva® than following the conventional method, which will facilitate In vitro fertilization and embryo transfer.

Application to valuable genetically modified mice:
Using CARD HyperOva® on your valuable genetically modified mice enables you to obtain a lot of ova from small number of females, thus enabling the large-scale preparation of embryos.

Periphere products

Additionally to reagents regarding reproductive engineering techniques we offer several instruments. For example a transportation kit for the cauda epididymides or straws to execute In vitro fertilisation steps.

Products

Media for In vitro Fertilisation

Name	Type	Productno.	Amount	References
For mouse
KSOM	In vitro fertilisation	CSR-R-B075 CSR-R-B074	10*5 ML 10*2 ML
mHTF	In vitro fertilisation	KYD-008-02-EX KYD-008-02-EX-X5 KYD-008-05-EX KYD-008-05-EX-X3	2 ML 5*2 ML 5 ML 3*5 ML	Kito et al., 2004
CARD MEDIUM®	In vitro fertilisation	KYD-003-EX	1 Kit	Takeo et al.,  2011 Takeo et al., 2010
FERTIUP® PM Card Medium Set	In vitro fertilisation	KYD-004-EX KYD-005-EX	1 Set 1 Set

Media for In vitro culture

Name	Typ	Artikelnr.	Menge	Referenz
For mouse
KSOM	In vitro culture	CSR-R-B075 CSR-R-B074	10*5 ML 10*2 ML	Lawitts et al., 1993
mWM	In vitro culture	CSR-R-B081 CSR-R-B080	10*5 ML 10*2 ML
0.25 M Sucrose	In vitro culture	CSR-R-Y078 CSR-R-Y077	10*5 ML 10*2 ML	Nakao et al., 1997
For rat
mR1ECM	In vitro culture	CSR-R-M174 CSR-R-M191	10*5 ML 10*2 ML

Media for preincubation of cryopreserved spermatocytes

Name	Typ	Artikelnr.	Menge	Referenz
FERTIUP® Preincubation Medium: PM	In vitro fertilisation	KYD-002-EX KYD-002-EX-X5 KYD-002-05-EX KYD-002-05-EX-X5	1 ML 5*1 ML 0.5 ML 5*0.5ML

Media for cryopreservation

Name	Typ	Artikelnr.	Menge	Referenz
FERTIUP® Cryoprotectant: CPA	Cryopreservation	KYD-001-EX KYD-001-EX-X5 KYD-001-05-EX KYD-001-05-EX-X5	1 ML 5*1 ML 0.5 ML 5*0.5 ML
1M DMSO	Cryopreservation	CSR-R-T072	10*2 ML	Nakao et al., 1997
DAP213	Cryopreservation	CSR-R-T073	10*1 ML

Media for embryo manipulation

Name	Typ	Artikelnr.	Menge	Referenz
PEPeS	Cryopreservation	CSR-R-P187	10*1 ML
P10	Cryopreservation	CSR-R-P186	10*2 ML
M2	In vitro manipulation	CSR-R-M084 CSR-R-M083	10*5 ML 10*2 ML
PB1	In vitro manipulation	CSR-R-P138 CSR-R-P185	10*5 ML 10*2 ML	Nakao et al., 1997

Media for an enhanced superovulation

Name	Typ	Artikelnr.	Menge	Referenz
CARD HyperOva®	In vitro fertilisation	KYD-010-06-EX KYD-010-06-EX-X5 KYD-010-EX KYD-010-EX-X5	5*1ML	Takeo et al., 2016 Takeo et al., 2015 Zitate: Noda et al., 2017 Nakagawa et al., 2016 Nakagawa et al., 2017

Disposable Items and instruments for murine reproductive engineering

Name	Typ	Artikelnr.	Menge	Referenz
Sperm Straws (10 pieces x 10 Units)	Disposable items, General labware	KYD-S020X10
Freezing Canister	Disposable items, General labware	KYD-S018	10 Units
Straw Connector (5 parts included)	Disposable items, General labware	KYD-S025	1 Unit
Triangular Cassette Short (10 units)	Disposable items, General labware	KYD-S021	10 Units
Triangular Cassette Long (10 units)	Disposable items, General labware	KYD-S035	10 Units
Embryo Manipulation Instrument Set	Disposable items, General labware	KYD-S036	1 Set
Glass capillary 20PC	Disposable items, General labware	KYD-S037	1 Set
Touch Burner APT-3	Disposable items, General labware	PHD-APT3-EX	1 Unit
CARD Cold Transport Kit	Disposable items, General labware	KYD-006-EX	1 Set