Renard J P, Babinet C
J Exp Zool. 1984 Jun;230(3):443-8. doi: 10.1002/jez.1402300313.
A method for obtaining a high survival rate of frozen-thawed mouse embryos is presented. Eight-cell mouse embryos were frozen inside small plastic straws in the presence of 1-2 propanediol and stored at -196 C. After thawing, the embryos were diluted for only 5 min in a 1.0 M sucrose solution to remove the 1-2 propanediol from the cells. At high rate of thawing (is equivalent to 2500 C/min) more than 88% of the embryos survived in vitro to the blastocyst stage provided that the dilution of propanediol was performed rapidly during thawing. At a lower rate of thawing (is equivalent to 300 C/min), survival tended to be higher (94.7%) when dilution was done 5 min after thawing. When the frozen-thawed embryos were transferred to the oviducts of day 1 pseudopregnant recipients either directly after the dilution of 1-2 propanediol or after 24 or 48 hr of culture, a high proportion of them (65.9%) develop normally to viable fetuses.
本文介绍了一种获得高冻融小鼠胚胎存活率的方法。将八细胞小鼠胚胎置于含有1,2 - 丙二醇的小塑料吸管中冷冻,并储存在-196℃。解冻后,胚胎在1.0M蔗糖溶液中仅稀释5分钟,以从细胞中去除1,2 - 丙二醇。在快速解冻速率(相当于2500℃/分钟)下,只要在解冻过程中快速进行丙二醇的稀释,超过88%的胚胎在体外存活至囊胚阶段。在较低的解冻速率(相当于300℃/分钟)下,解冻后5分钟进行稀释时,存活率往往更高(94.7%)。当冻融胚胎在1,2 - 丙二醇稀释后直接或在培养24或48小时后转移到第1天假孕受体的输卵管中时,其中很大一部分(65.9%)能正常发育成有活力的胎儿。