Traditional Cryopreservation is done using tranditional mating to generate 8-cell mouse embryos derived from 2.5 dcp (days post-coitum) females for freezing. Speed Cryopreservation is done using in vitro fertilization to generate 2-cell embryos for freezing. Embryos are stored in liquid nitrogen to provide a long term source of the line.
•To permanently store mouse lines.
For traditional cryopreservation, timed pregnant females (up to 3 session) will be obtained and sacrificed at 2.5 dpc. Embryos will be collected from the oviduct, and aliquots of 10-20 embryos will be frozen in a cryovial. The investigator should be aware that traditional cryopreservation often requires an expansion of the colony to obtain sufficient numbers of male and female mice of the appropriate age for generating the timed pregnant females. Individual male mice are needed for these matings and this will also lead to increased housing costs.
Speed cryopreservation uses in vitro fertilization to generate the embryos for cryopreservation. Sperm from the mouse line to be preserved are used to fertilize eggs generated from female wild type or colony mice. Female mice will be superovulated and oocytes will be collected. Sperm from a male mouse that is a proven breeded will be collected and used to fertilize the oocytes. After incubation, 2-cell embryos will be collected and frozen in cryovials in aliquots of 10-20 embryos. The advantage of this method is that the colony does not have to be expanded to accomodate this approach. Female wild type mice can be purchased at the right age from an external or in-house vendor and only the male mouse needs to come from the colony.
The frozen embryos will be stored in the Facility, unless a different storage location is requested. The cost of storage is currently $150 per year for up to 6 lines. The animal protocol for the mice and procedure is covered by the Facility. The investigator should be aware that the number of embryos that can be collected from these methods varies significantly among strains and although the facility attempts to store at least 100 embryos per line, this cannot be guaranteed.
The Investigator Must Supply
• Female or male mice from which to generate the embryos
• Housing of mice during breeding for generation of embryos.
$1000 for traditional embryo cryopreservation for up to three sessions. Additional charges may apply if more sessions are required to obtain suitable numbers of embryos for storage.
$800 for speed cryopreservation of embryos (Note that the cost of wild type females for superovulation is not included in this price).
$1000 for speed cryopreservation of embryos and sperm cryopreservation. THIS IS OUR RECOMMENDED OPTION!
•Storage in liquid nitrogen (Fee $150.00/yr for up to 6 lines)
•Embryo Transfer into CD1 pseudo-pregnant female (Fee - $250.00/attempt)
•Embryo Culture to blastocyst stage (Fee - $50.00)
Enquiries and requests for services should be made though our on-line Requisiton of Services