冻融兔卵母细胞培养不同时间再激活效应的研究

The effect of activation on the development potential of frozen-thawed rabbit oocytes cultured for different time after fertilized by ICSI

目的探讨冻融兔卵母细胞行卵胞浆内单精子注射(ICSI)后,不同培养时间、不同激活方法,卵母细胞受精及胚胎发育能力。方法共收集兔卵母细胞470枚,玻璃化冷冻,解冻行ICSI后培养半小时,分为3组激活:组1(n=46):钙离子激活;组2(n=40):氯化锶(SrCl_2)激活;组3(n=33):无水酒精激活;再同样方法解冻一部分卵母细胞,ICSI后培养1 h,分为2组激活:组4(n=30):钙离子激活;组5(n=26):氯化锶激活;对照组(n=39):ICSI后直接培养,不激活。各组得到的囊胚玻璃化冷冻,受体兔HCG准备后,胚胎解冻后移植。结果卵母细胞解冻行ICSI后,组5的卵母细胞受精率、分裂率和囊胚形成率最高,分别为53.8%,26.9%和7.7%,所获囊胚的母兔可能怀孕。结论冷冻卵母细胞解冻行ICSI后,适当延长培养时间,使用合适的激活方法可能会提高卵母细胞的受精以及早期胚胎发育能力。

Objective. To investigate the effect of activation on the development potential of the fertilized frozenthawed rabbit oocytes cultured for different time after fertilization. Methods. Rabbit oocytes were vitrified. After thawing, the oocytes were fertilized by intracytoplasmic sperm injection （ICSI） and cultured for half an hour and then divided into three groups at random. Group 1 （n=46）. oocytes were activated by calcium ionophore; Group 2 （n=40）. activated by strontium chloride; Group 3 （n=33）; activated by anhydrous alcohol. Another part of frozen-thawed oocytes were fertilized by ICSI and cultured for an hour and then divided into three groups at random. Group 4 （n=30）; oocytes were activated by calcium ionophore; Group 5 （n=26）; activated by strontium chloride; Control group （n= 39）. without activation. Blastocysts derived from each group were transferred to recipient rabbits. Results.. Rates of fertilization, cleavage and blastocyst formation of Group 5 were the highest. The rabbits transferred with the embryos derived from Group 5 may become pregnant. Conclusions： Suitable activation and reasonably longer culture time after frozen-thawed oocytes fertilized by ICSI may improve the oocyte developmental potential.