Mouse Line Resuscitation-Rederivation
Resources and recommendations
The Department of Comparative Medicine has several quarantine rooms available to research staff who require the importation of animals from unapproved vendors. If necessary, the TMM will work with DCM to perform rederivation of “dirty” animals. Contaminated animals will be mated, 2-8 cell embryos will be subjected to a washing procedure and transferred into pathogen-free, (SPF) foster mothers. Foster mothers will be housed in SPF conditions in a special clean quarantine room under the supervision of TMM and DCM personnel. At 14 days of age DCM personnel will collect samples and send them to a diagnostic laboratory for evaluation. Following confirmation of pathogen-free health status, the animals will be transferred to the rodent facility of your choice for housing and use.
Responsible parties and timeline
Step | Timeframe | Protocol | Responsible party |
---|---|---|---|
1 | Week 1 | Provision of dirty mutant stud males | Customer |
2 | Week 2 | Embryo generation and transfer to foster mom | TMM |
3 | Week 7 | Biopsy collection and transfer to Charles River lab | DCM |
4 | Week 8 | Transfer clean offspring to customer | DCM |
5 | Variable | Genotyping of pups | Customer |
Service Ordering for Rederivation
To initiate a rederivation project, fill out and forward a Resuscitation/Rederivation Request Form to TMM or contact us via email or phone. There will be an additional form from DCM to reserve space in the SPF quarantine room to fill out before we begin your project.
In addition to the Project Request Form, you will need to provide the following materials:
a) 2-3 mutant stud males from the line you are importing.
b) 5-6, 3-4 week old mutant females from the line you are rederiving (if you want to have homozygous mouse stock, otherwise we can supply C57 BL/6NJ females)
Resuscitation of mouse lines using frozen germplasm (embryos or sperm)
Germplasm is very sensitive to temperature changes therefore it is delivered in a Dry shipper cooled by LN2.
Rederivation Service Ordering
- To initiate the project, please fill out and send a Project Request Form to the TMM.
- Once you have arranged an MTA with the sending partner please place us in contact with them to make shipping arrangements. During this process we please ask that you request two samples as an insurance policy should the first prove unsuccessful. We have a dry shipper available for rental to help expedite the process.
- Once we receive the samples we will transfer them into our LN2 dewer for storage until we can schedule your procedure.
Standard rederivation service fees include
- Purchase of 3 week old C57BL6/NJ or B6xDBA-2F1 females (donors of oocytes).
- Superovulation of 3 week old females.
- Collection and Transplantation of rederived embryos into pseudopregnant foster females.
- The production of guaranteed clean animals.
- Animal housing charges for fosters and their offspring until weaning.
Service fees do not include
- Ordering of special strains of female mice for oocyte donors (see additional services below).
- Animal housing charges for litters after weaning.
- Ear/tail tissue collection for DNA isolation for genotyping.
- Health testing of the foster mother and pups.
- Shipping charges.
Additional services
At your request TMM can purchase and use other inbred mouse lines for the rederivation of mutant mice. Strains that cost significantly more than C57 BL/6NJ will be charged an additional fee based on the difference in cost.