Sperm Resuscitation Service
For most mouse strains, we recover cryopreserved sperm by IVF. For sperm with zona-penetration defect or other infertility phenotypes caused by gene modifications, we will recover the sperm using laser-assisted IVF (Li et al. 2013) or ICSI method (Li and Lloyd, 2006).
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.
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 cryopreservation 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 7.5 IU eCG by followed by IP injection of 7.5 IU hCG 47 hours later.
- If sperm from a mouse line have sperm-cumulus penetration defect, cumulus-free IVF can be performed using oocytes with their cumulus layers removed by treatment with hyaluronidase.
- If sperm from a mouse line have sperm-zona binding or sperm-zona penetration defect, laser-assisted IVF can be performed. Laser-zona drilling (LZD) has been recognized for being a safe and effective method to assist IVF in mice.
Please see our order page for additional information on IVF pricing.
Li MW, Kinchen KL, Vallelunga JM, Young DL, Wright KD, Gorano LN, Wasson K, Lloyd KC 2013 Safety, efficacy and efficiency of laser-assisted IVF in subfertile mutant mouse strains. Reproduction 145 245-254.
Li MW, Lloyd KCK 2006 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, p23-40.
Li MW, and Lloyd KCK et al. (2013) Safety, efficacy and efficiency of laser-assisted IVF in subfertile mutant mouse strains. Reproduction 145: 245-54.
Li MW, KCK Lloyd et al. (2014) IVF recovery of mutant mouse strains using sperm cryopreserved with MTG in cryovials. CyroLetters 35: 145-53.
Li MW, Lloyd KCK (2006) 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, p23-40.