CARD MEDIUM®

Kyudo

Catalog No.: KYD-003-EX

Refer to the Reproductive Engineering Techniques in Mice Technical Manual for complete instructions of IVF

Product Description

CARD MEDIUM® is a novel culture medium that acts during IVF on the mouse egg zona pellucida to promote high fertilization rates, especially when used in conjunction with CARD FERTIUP® Preincubation Medium: PM. CARD MEDIUM® was developed by The Center for Animal Resources and Development (CARD), Kumamoto University, Japan.

CARD MEDIUM® is an IVF culture medium that acts on the zona pellucida of the egg during IVF and improves the fertilization rate. Combined use of FERTIUP® sperm cryoprotectant and CARD FERTIUP® Preincubation Medium: PM greatly improves the fertilization rate of frozen-thawed sperm in C57BL/6 and 129 strains, from less than 10% using conventional methods to more than 30%. Further, when coupled with our new CARD MEDIUM®, fertility rates are boosted to over 80%. Thus, combined usage of the trio of CARD FERTIUP®Cryoprotectant: CPA, CARD FERTIUP® Preincubation Medium: PM and CARD MEDIUM® provides a high and consistent rate of fertilization from frozen-thawed mouse sperm, which will allow you to efficiently recover mouse lines when needed.

For information on techniques for mouse spermatozoa cryopreservation, spermatozoa thawing, in vitro fertilization and obtaining pups through embryo transfer, please refer to the CARD website: https://card.medic.kumamoto-u.ac.jp/card/english/sigen/manual/onlinemanual.html

CARD MEDIUM® FAQs

Can I achieve high IVF rates without using the CARD FERTIUP series together with CARD MEDIUM® (Cat. No. KYD-003-EX)?
The CARD FERTIUP series acts on sperm, while CARD MEDIUM® acts on oocytes to improve fertilization rates. Therefore, using CARD MEDIUM® together with the CARD FERTIUP series can achieve IVF rates of over 80% even in C57BL/6 mice.

Can I achieve high IVF rates in mouse strains other than C57BL/6 using CARD MEDIUM® (Cat. No. KYD-003-EX)?
Yes. High IVF rates have been achieved in a variety of mouse strains when CARD MEDIUM® is used in combination with the CARD FERTIUP series.

Reference:
Immunotherapy using inhibin antiserum enhanced the efficacy of equine chorionic gonadotropin on superovulation in major inbred and outbred mice strains
Takeo T, Nakagata N. Theriogenology. 2016;86(5):1341-1346. doi:10.1016/j.theriogenology.2016.04.076.

I found that many sperm were attached to oocytes when performing IVF using CARD MEDIUM® (Cat. No. KYD-003-EX) with fresh mouse sperm. Is there a risk of polyspermy?
Mouse oocytes have two mechanisms that prevent multiple sperm from entering the cytoplasm. Even when many sperm attach to the zona pellucida surrounding the oocyte, the zona reaction prevents additional sperm from penetrating the zona pellucida. Furthermore, even if a sperm passes through the zona pellucida, the oocyte membrane provides an additional block to sperm entry. Therefore, it is very rare for more than one sperm to enter the oocyte cytoplasm (polyspermy). In particular, the incidence of polyspermy in C57BL/6 mice is extremely low, approximately 0.2–0.3%, and is not considered a concern.

I performed IVF using CARD MEDIUM® (Cat. No. KYD-003-EX), but embryo development did not progress properly beyond the 2-cell stage. The embryos were cultured in KSOM medium or Whitten medium. What could be the cause?
Several factors may contribute to poor embryo development:
1. The incubator temperature or CO₂ concentration may not be optimal.
2. If a homemade embryo culture medium is being used, the distilled water or medium components may be affecting embryo development. Consider using a commercially available medium such as CARD KSOM (Cat. No. KYD-011-EX).
3. If the mineral oil overlaid on the culture medium is old, embryo development may be impaired.