In Vitro Fertilization (IVF)

1. Oocytes: Labs need to provide 8-12 young (3-5 wks old) females for superovulation. These can be colony raised or purchased through vendors. All females must be delivered to the GEAM’s rederivation rooms at MSC (rats room 3045, mice room 3020) and GEAM will superovulate them.
2. Fresh sperm: For lines that need rescuing, one male breeder (preferred) is delivered to GEAM on the morning of the IVF or the day prior. GEAM will sacrifice him and assess sperm quality. If motility is >10%, sperm is used for IVF and excess cryopreserved if the lab requests.
3. Frozen sperm: Sperm will be thawed via instructions from the repository. If the instructions suggest using media not typically used, GEAM will make the solution and charge the investigator ($125/hr). Frozen samples are thawed on the morning of the IVF.
4. Co-incubation:  Oocytes are co-incubated with sperm (whether fresh or frozen) for five hours. After that co-incubation, presumptive zygotes are washed through pre-equilibrated media and incubated overnight to access embryonic development to two-cell stage.
5. Embryo transfers: The next morning, two-cell embryos will be transferred into psuedopregnant females to recover live offspring.

Disclaimer: The success of the IVF is dependent on the mouse strain, sperm motility, sperm counts, post-thaw motility and egg quality.