Cryopreservation of Rat Blastocysts by Vitrification Using Ethylene Glycol, Polyethylene Glycol and Sucro

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Rat blastocysts were cryopreserved by vitrification with a simple and rapid method using ethylene glycol (EG)-based solution which was mixed with different concentrations of polyethylene glyco1 (PEG)(5-15%) and sucrose (0.25-1.OM). Effect of time (2-10min) and temperature (on ice or at room temperature) for equilibration on the in vitro development of vitrified-warmed embryos were investigated.Successful vitrification was obtained by the equilibration for 5 min on ice (3-4C). More than eighty percent of the vitrified-warmed embryos developed to the stages of expanded or hatched blastocyst when the embryos were equilibrated on ice for 5 min in the solution containing40%EG, 5%PEG and 0.5M sucrose.The solution containing sucrose (0.5M) was not effective for recovering the vitrified-warmed embryos. About 30% of the vitrified-warmed embryos developed to live fetus after transfer to recipient female rats. The present results indicate that even a simple and rapid method could be successful for the vitrification of rat blastocysts using the solution containing EG, PEG and sucrose.
日本哺乳動物卵子学会の論文