Resuscitation

The MBP has over 20 years of experience collecting and cryopreserving mouse germplasm, which includes an established program for cryorecovering mouse strains. Each year we successfully perform an average of 250 reanimation procedures from preserved mouse sperm or embryos for researchers around the globe.

Sperm Resuscitation Service

For most mouse strains, we recover cryopreserved sperm by IVF. For sperm with extremely low sperm quality or infertility phenotypes caused by gene modifications, we will recover the sperm by ICSI.

Embryo Resuscitation Service

For embryo resuscitation, we thaw cryopreserved embryos and transfer them into oviducts (2-cell stage) or uteruses (morula and blastocyst stages) of pseudopregnant recipients, and then let the embryos develop to pups in vivo.

Sperm and embryos for resuscitation can be provided by you or can be obtained from an outside source. Our importation staff can assist you on various levels with the importation of outside materials. Please include a thawing protocol (sperm or embryo) and information on embryo stage at cryopreservation. It is recommended that two vials or straws per strain are shipped to us in a liquid nitrogen dry shipper.

Please see our order page for additional information on cryorecovery pricing.

In Vitro Fertilization (IVF)

In vitro fertilization (IVF) is a process by which superovulated mature eggs are fertilized by sperm, in vitro, after sperm capacitation. IVF is used for embryo cryopreservation, sperm resuscitation, rederiving a “dirty” mouse strain to Specific Pathogen free (SPF) barrier facility, speed expansion, and to rescue a line if males are poor breeders or die unexpectedly.

Sperm are collected from cauda epididymides and preincubated for capacitation before they are used for insemination. Cumulus-oocyte complexes (COCs) are collected from the oviducts of superovulated females 14-16 hours post hCG injection. Superovulation is induced by IP injection of 5-7.5 IU eCG by followed by IP injection of 5-7.5 IU hCG 47 hours later.

Please see our order page for additional information on IVF pricing.

References:

Li MW, Lloyd KCK. DNA fragmentation index (DFI) as a measure of sperm quality and fertility in mice. Sci Rep. 2020; 10(1):3833. doi: 10.1038/s41598-020-60876-9.

Li MW, Glass OC, Zarrabi J, Baker LN, Lloyd KC. Cryorecovery of mouse sperm by different IVF methods using MBCD and GSH. JFIV Reprod Med Genet 2016; 4, 175; doi: 10.4172/2375-4508.1000175

Li MW, Vallelunga JM, Kinchen KL, Rink KL, Zarrabi J, Lloyd KC. 2014. IVF recovery of mutant mouse strains using sperm cryopreserved with MTG in cryovials. Cryo Letters. 35:145-153.

Li MW, Kinchen KL, Vallelunga JM, Young DL, Wright KD, Gorano LN, Wasson K, Lloyd KCK. Safety, efficacy and efficiency of laser-assisted IVF in subfertile mutant mouse strains. Reproduction. 2013; 145:245-254.

Li MW, Lloyd KCK. Intracytoplasmic sperm injection (ICSI) in the mouse. In “Principles and Practice: Mammalian and Avian Transgenesis – New Approaches”, Shirley Pease and Carlos Lois (Eds), Springer–Verlag Berlin Heidelberg, 2006; p23-40.