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 cells embryos will be subjected to a washing procedure and transferred into pathogen-free, (SPF) foster mothers. Foster mothers will be housed in SPF isolators in special clean quarantine rooms under control of TMM and DCM personnel. After the pups are weaned 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 for housing and use.
Responsible parties and timeline
|1||Week 1||Provision of dirty mutant stud males||Customer|
|2||Week 2||Embryo generation and transfer to foster mom||TMM|
|3||Week 5||Biopsy collection and transfer to Charles River lab||DCM|
|4||Week 7||Transfer clean offspring to customer||DCM|
|5||Variable||Genotyping of pups||Customer|
To initiate a rederivation project, fill out and forward a Resuscitation/Rederivation Request Form to TMM or contact them via email or phone.
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) 2-3 virgin mutant females from the line you are importing (if you want to have homozygous mouse stock)
Resuscitation/Rederivation of mouse lines using frozen germplasm (embryos or sperm) delivered from outside of OHSU
Germplasm is very sensitive to temperature changes therefore it is delivered in Dry shipper with temperature of LN2.
- To initiate the project, please fill out and forward a Resuscitation/Rederivation Request Form to the TMM.
- When you know suspected delivery date inform TMM Personnel several days in advance about shipment by mail and phone talk.
- Please order the shipment of Dry shipper with embryos or sperm from your partner directly to address TMM. Shipment must be done on Monday or Tuesday to be sure that Dry shipper will be delivered during working week to TMM.
- Inform TMM about the shipment and delivery date 3-4 days in advance that TMM will be ready to accept your and save germplasm correctly. Alternatively TMM will not be responsible for germplasm quality. It will be nice if your partner will inform you and TMM about shipment simultaneously
- Do not open Dry shipper by yourself! TMM personnel will open Dry shipper by themselves. Alternatively TMM will not be responsible for germplasm quality.
Standard rederivation service fees include
- Purchase of virgin C57BL6/J or B6xDBA-2F1 females (donors of oocytes).
- Superovulation of virgin females.
- Collection and Transplantation of rederived embryos into pseudopregnant recipients.
- The production of guaranteed clean animals.
- Animal housing charges for recipients and their offspring until weaning.
Service fees do not include
- Animal housing charges for litters after weaning.
- Ear/tail tissue collection for DNA isolation for genotyping.
- Health testing of the foster mother.
- Shipping charges.
According to customer’s request TMM can purchase and use other inbred mouse lines for the rederivation of mutant mice. Moreover, TMM can tag, collect tail samples, extract DNA and genotype the offspring by PCR. For this purpose the animals can be housed in our colony beyond the usual 3 weeks (weaning age). These additional services will be charged additional fees including extra costs for the cage per diem.