Cryopreservation and Recovery

Fees for Service

The UTSW TTC will cryopreserve fertilized eggs, sperm and ovaries from transgenic and knock-out strains of mice. The Center will store the cryopreserved gametes. The Center will, on request, thaw the cryopreserved bisected ovaries and surgically transfer the ovary to histocompatible recipients. The Center will, on request,  recover the cryopreserved embryos and transfer them to pseudo-pregnant recipients.  The Center will, on request, thaw the cryopreserved sperm, perform in vitro fertilization, culture the eggs to the 2-cell stage and transfer the embryos to pseudopregnant recipients.

                                            Cryopreservation – procedure                             

                                                        $600 – embryos
                                                        

                                                        $500 – ovaries

                                                        $500 – sperm

                                                Cryopreservation – storage    $100/year


                                           Embryo recovery and transfer     $600 – 2 to 4 transfers


                                 Ovary recovery and transplantation      $1,000 – 2 to 4 transfers

Sperm recovery, in vitro fertilization & embryo transfer     $2,000 – 2 to 4 transfers

Guarantee Policy

In exchange for the fees charged for services, the Center warrants that it will cryopreserve ~200 embryos or 6 ovaries or 10 straws of sperm from transgenic/knockout mice. The embryos will be stored in cryovials at ~35/vial.  10 to 14 embryos will be stored in a vial for quality assurance. The bissected ovaries will be stored 1/2 ovary/vial.  The sperm will be stored in straws at approximately 10µl/straw, 10 straws/male.

Investigator Requirements

1.     Investigator must submit the Animal Reassignment Request Form for females carrying transferred eggs or ovaries to be transferred and housed in the ARC quarantine.

2.     The Investigator contacts Mylinh Nguyen (x56287) to discuss and schedule the cryopreservation or the recovery processes.

3.     The Investigator submits the appropriate Cryopreservation or Recovery Request Forms for services.

4.     For embryo cryopreservation, the Investigator will transfer to the TTC,  7 to 8 proven stud males between 10 to 16 weeks of age.  The Center will not accept unproven males or senescent males for this service.  The males birthdate, genetic background and genotype must be provided.  Following the cryopreservation, the males can be returned to the Investigator’s colony but they will not be considered SPF animals.

5.     The TTC staff will perform the hormone priming of WT females (B6, 129SvEv(S6), B6SJLF1), and set up the appropriate matings.  If the Investigator prefers to cryopreserve the line in a homozygous condition, the Investigator will need to supply the females of the appropriate number and age to perform the procedure.  Please consult with the TTC staff to arrange this alternative.

6.     For sperm cryopreservation, the Investigator will transfer to the TTC, 2 or 3 stud males between 10 to 16 weeks of age.  The Center will not accept unproven males or senescent males for this service.  The males birthdate, genetic background and genotype must be provided.

7.     For ovary cryopreservation, the Investigator will transfer to the TTC, 2 fertile females between 4-10 weeks of age.  The females birthdate, genetic background and genotype must be provided.

8.     For ovarian transplantation, the TTC staff will surgically implant thawed, bisected ovaries (2-4) into the ovarian bursa of female recipients of the appropriate genetic background.  The TTC will purchase female recipients and perform the transplantation when they are approximately 5-6 weeks of age.

9.    For embryo recovery and transfer, the Investigator provides the TTC staff with the frozen vial or straw of embryos and a protocol from the source of the embryos detailing the thaw methodology.  The Investigator should provide from 25 to 50 embryos to ensure successful recovery of the line.

10.  For sperm recovery, IVF and embryo transfer, the Investigator supplies the TTC with the straws of cryopreserved sperm that have been shipped to the TTC via a liquid nitrogen dry shipper.  The Investigator should provide the TTC with the name of the line, genetic background and genotype.  The TTC will perform the IVF using wild-type C57Bl/6 eggs unless otherwise specified.  The Investigator must provide a protocol from the original source of the sperm that details the methodology used for the successful recovery of the sperm.