乙二醇玻璃化冷冻液二步法对长爪沙鼠2-细胞胚胎的冷冻保存

Cryopreservation of Mongolian gerbil 2-cell Embryos by Vitrification based on EG with Two Steps

目的评价用乙二醇玻璃化冷冻液二步法对长爪沙鼠2-细胞胚胎冷冻保存的效果。方法通过超排卵处理后与雄鼠交配获得长爪沙鼠2-细胞胚胎,胚胎在冷冻液EFS20中平衡3 min,然后移入冷冻液EFS40中平衡1 min,之后将细胞冻存管直接浸入液氮中保存。采用二步法复苏胚胎,复苏后移入改良的胚胎培养液(mKSOM)中体外培养。结果采用该方法冷冻的胚胎复苏后胚胎回收率为86.73%,存活率为82.94%,复苏后存活的胚胎中约1/3胚胎可以在体外发育到囊胚。结论乙二醇玻璃化冷冻液适于冷冻保存长爪沙鼠2-细胞胚胎。

Objective Using the thyleneglycol（EG）-based solutions with two steps to freeze the 2-cell embryos of Mongolian gerbil（Meriones unguiculatus） and to appraise the effect of embryo cryopreservation. Method The embryos were exposed serially to EFS20 for 3 minutes and EFS40 for 1 minute. The embryo trealed were immersed into liquid nitrogen. Result The embryo recovery rate was 86.73% and after freezing-thawing it survival rate was 82.94%. And about 30% of the freezing-thawing embryos developed to blastocyst in vitro. Conclusion This results demonstrates that Mongolian gerbil embryos can be safely eryopreserved using vitrification solutions based on EG with two steps.