| Rapid cryopreservative and vitrified methods were used to freeze the mouse morulae and blastocyst, to explore different concentrations of ethylene glycol (EG) and EFS series on the impact of embryonic development and to research the feasibility and the development rate by double cryopreservation.The results show that:1. By the test of vitrification and devitrification , EFS20, EFS30 and EFS40 can be used as a mouse embryo vitrification solution.2. The study of toxicities of the vitrification solutions showed that when embryos were e quilibrated in mPBS , development in vitro was not effected;low concentrations of ethylene glycol (1.8 mol / L ~ 3.6mol / L) almost has non-toxic, however, high concentration of ethylene glycol (4.5 mol / L) has a certain toxicity ;in EFS20, EFS30 and EFS40 in a short processing time (0.5 min) almost has non-toxic, but with the increased concentration and the processing time extension, toxicity is also growing.3.By slow freezing with different concentrations of ethylene glycol and vitrification with different concentrations of EFS series, the development rate of morulae is 52.8% ~92.5% and 78.9% ~ 91.5%, the development rate of blastocyst is 38.6%~82.8% and 72.5%~81.3%. The best frozen effect is 1.8mol/LEG and EFS30 (morulae was 92.5% and 91.5%, the blastocyst was 82.8% and 81.3%).4. The method of slow freezing was not significantly different from the method of vitrification .5. The freezing tolerancet of morulae was better than blastocyst.6. By the 1.8 mol / LEG slow freezing and double cryopreservation with 1.8 mol / LEG and EFS30 the development rate of morulae was 46.7% and 53.8%, the blastocyst was 33.9% and 26.4%; by the vitrification with EFS30 and double cryopreservation with 1.8 mol / LEG and EFS30 the development rate of morulae was 39.4% and 41.3%,the blastocyst was 29.1% and 31.0%. Double cryopreservation is feasible to mice embryos, it is alse applied as examining approaches in freezing tolerance,freezing method and cryoprotective agent.
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