The Transgenic Facility provides rederivation services to all AECOM investigators seeking to maintain or save cell lines/strains. We advise individual labs to refrain from waiting until all candidate mice are over the age of 4 months, as advanced age decreases the chances of rederivation success. Investigators must fill out a Rederivation Form and contact the Transgenic Facility Director to schedule the procedure and discuss the details of candidate cell lines/strains. Scheduling is on a first come-first served basis.
Individual investigators are responsible for the super ovulation and mating of candidate mice. once a target date has been agreed upon, investigators must obtain super ovulation hormones and a tube of embryo-specific media from the facility. Information on super ovulation scheduling can be found here. Training for either plug-checking or oviduct isolation can also be arranged with the facility director. The super ovulation hormones used are Pregnant Mare Serum (PMS) and hydrolyzed chorionic Gonadotropin (HCG). PMS is distributed in the form of a frozen 50 IU microcentrifuge tube. HCG is distributed in the form of a 50 IU lyophilized powder in a microcentrifuge tube. Keep in mind when scheduling the injections that you must give the first injection (PMS) three days before the agreed upon target date for the rederivation procedure.
Immediately after HCG injection, set up your mating, preferably a single cage with one hormonally treated female with one male. It is recommended you set up a maximum of two females for every male. The following morning, check to see if any females are plugged and separate any unplugged females from the males. If any super ovulated female has not plugged, they will not be able to plug for at least 5-7 days, therefore it is advised that they be removed and set aside to recover. Isolate the oviducts of all plugged females and place dissected oviducts into a Petri dish containing embryo-specific media (M2, FHM). Be very careful to not touch the dish with anything that touched the animals. Change gloves frequently during the procedure. Wipe the dish with 10% bleach while being careful to not use excessive bleach so as to compromise the embryos in the dish.
Assign a member of your lab, that has not had any contact with animals or been in an animal room, to take the dish directly to Price B110. No one who has had contact with animals or has been in any animal room within the past 24 hours is allowed into the facility labs in Price B110.
Facility personnel will then dissect the oviducts, isolate the embryos and transfer them into a CD1 foster mother. After the embryo transfer is complete the facility will call your lab to pick up the cage(s) of foster mothers. We cannot house any cages for investigators once the rederivation is complete. It is the responsibility of individual investigators to secure space in an animal room, via Institute of Animal Studies, before requesting a rederivation. In 19-20 days, foster mothers should give birth. Please contact the facility and inform us of the number of pups born.