影响玻璃化冷冻兔胚胎效果的一些因素

试验对影响玻璃化冷冻兔胚胎效果的一些因素进行探讨，以找出理想的玻璃化冷冻方法。在测试的５种玻璃化溶液中，含３５％乙二醇（ＥＧ）和１．０ｍｏｌ／Ｌ蔗糖的溶液（ＶＳ１）对胚胎的毒性最小。用ＶＳ１冷冻桑椹胚和囊胚的理想程序是：在室温下使胚胎分别在２０％ＥＧ和３５％ＥＧ中平衡２、３分钟后，移入ＶＳ１中，０．５分钟内（囊胚也可在２分钟后）投入液氮中冷冻。桑椹胚的存活率为９１．７％（３３／３６），囊胚的存活率为９７．１％（３３／３４）～９７．３％（３６／３７）。８～１６细胞胚胎的理想冷冻程序为：在室温下使胚胎在２０％ＥＧ、３５％ＥＧ中平衡２、３分钟，移入４℃的３７％ＥＧ＋１．０ｍｏｌ／Ｌ蔗糖溶液中平衡２分或１０分钟后冷冻，胚胎存活率分别为１００％（３７／３７）、８６．１％（３１／３６）。

Some Factors Affecting the Survival of Rabbit Embryos Vitrified in Ethylene Glycol Sucrose Solution

Four experiments were conducted to examine the factors affecting the survival of vitrified rabbit embryos and establish the optimum vitrification procedures.The results of toxicity test demonstrated that the solution consisting of 35% ethylene glycol(EG) and 1.0M sucrose in PBS(VS 1) was the least toxic among the 5 vitrification solutions tested.Results of vitrification cxperiments showed that the optimum procedures for vitrifing morula and blastocyst were to cxpose the embryos to 20% EG、35%EG at 25℃ for 2and 3 min respectively followed by exposure to VS 1 for less than 0.5 min or for 2 min(for blastocyst only)and vitrification in liquid nitrogen.The survival rate of embryos vitrified using these procedures was 91.7%(33/36) for morula and 97.1%(33/34)～97.3%(36/37) for blastocyst.The optimum procedures for the vitrification of 8 to 16 cell embryos were to expose the embryos to 20%EG、35%EG at 25℃ for 2 and 3 min respectively,followed by exposure to 37% EG+1.0M sucrose at 4℃ for 2 to 10 min and vitrification.The survival rate of vitrified 8 to 16 cell embryos was 86.1%(31/36)～100%(37/37).