RRC ID 28834
Author Taketsuru H, Kaneko T.
Title Efficient collection and cryopreservation of embryos in F344 strain inbred rats.
Journal Cryobiology
Abstract In rats, it is now possible to produce genetically engineered strains, not only as transgenic animals but also using gene knockout techniques. Reproductive technologies have been used as indispensable tools to produce and maintain these novel valuable strains. Although studies for collecting and cryopreserving embryos have been reported using outbred rats, efficient methods have not been established in inbred strains. The F344 inbred strain is important in rat breeding and has been used for the production of transgenic/knockout strains and for genome sequencing. Here we studied the optimal conditions for oocyte collection by induction of superovulation, and the development of embryos after cryopreservation in F344 rats. The response to pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) was examined by injection of 150 IU/kg PMSG + 75 IU/kg hCG or 300 IU/kg PMSG + 300 IU/kg hCG. Superovulation was achieved at high efficiency by an injection of 150 IU/kg PMSG + 75 IU/kg hCG. Furthermore, superovulation in this strain showed similar high response as Wistar rats. Of 2-cell embryos cryopreserved by vitrification in a solution containing 10% propylene glycol, 30% ethylene glycol, 20% Percoll and 0.3 M sucrose, more than 90% survived after warming and 32% developed to offspring. However, the freezability of pronuclear stage embryos was extremely low. This study demonstrated that sufficient unfertilized oocytes and embryos can be collected from F344 rats by the induction of superovulation with 150 IU/kg PMSG + 75 IU/kg hCG. Furthermore, cryopreservation of 2-cell embryos using this vitrification protocol can now be applied to maintaining valuable rat strains derived from the F344 inbred strain as genetic resources.
Volume 67(2)
Pages 230-4
Published 2013-10-1
DOI 10.1016/j.cryobiol.2013.07.004
PII S0011-2240(13)00195-8
PMID 23928269
MeSH Animals Chorionic Gonadotropin / pharmacology Cryopreservation / methods* Cryoprotective Agents / chemistry Embryo, Mammalian / physiology* Female Gonadotropins, Equine / pharmacology Horses Humans Oocytes / physiology Ovulation Induction / methods Pregnancy Rats / embryology* Rats, Inbred F344 Rats, Wistar Superovulation / drug effects Vitrification
IF 2.287
Times Cited 10
Rats F344/Stm(strainID=266) LE/Stm(strainID=265)