IVF service is used to rederive a mouse line imported from other institute, to rescue a mouse line that is in danger of being lost, to resuscitate a mouse line from previously cryopreserved sperm, or to facilitate mating for backcross/production. Live mice housed at BCM or cryopreserved sperm can be used as a source material.
What Will Happen
- The Investigator will submit an IVF request form through iLab with detailed information about the sperm or mice. If cryopreserved sperm from an outside source is to be used, please have the IVF request in place prior to delivery of the sperm. the IACUC protocol number and account must be provided at this time. The Investigator’s IACUC protocol must be approved for all procedures performed in the Investigator’s laboratory.
- Sperm or live mice delivery to be coordinated by the investigator.
- a. If live mice are used, please submit a request to have the male mouse (mice) transferred by CCM or delivered by the Investigator to ABBR R119 by 8 am on the scheduled IVF day. For CCM transfers, please ark clearly on the transfer request that the mice are for IVF, must be transferred the same day as the scheduled request for IVF, and are required to be in place by 8 am. The core will recover sperm from the male for the IVF procedure. The male(s) will be euthanized and cannot be returned to the Investigator.
- b. If cryopreserved sperm is used, the core will schedule the IVF procedure after the sperm arrives. The investigator can have the sperm shipped to their own lab and hand-deliver to the GERM Core or they can have the sperm shipped directly to the core with prior communication.
- The core will perform IVF on wild-type oocytes from C57BL/6J, C57BL/6N, FVB or ICR mice. 15 female mice will be used for the IVF with superovulation. 300-500 oocytes are expected to be able to collect for the IVF.
- After overnight culture, all 2-cell embryos will be transferred to ICR pseudo pregnant females to carry gestation to term in core.
- Live-born founder animals will be held by the GERM Core until 10-14 days of age and subsequently transferred to the Investigator. Investigators will perform genotyping to identify the desired founder mice.
What is Expected
- The number of pups to be generated by IVF varies widely, from less than 10 to over 60 pups, with occasionally no pups born. Sperm quality directly correlates with the number of oocytes able to be fertilized, and subsequently the number of pups born.
- If no pups are born, the core offers to repeat the IVF at 50% cost if there is another male mouse or straw of sperm available.
- Please contact the core if use of inbred strains other than C57BL/6J, C57BL/6N, FVB, or ICR are desired.