两种激活方案对辅助生殖技术中未受精卵母细胞的激活效果比较

Comparison of Two Methods on Activation of Human Unfertilized Oocytes after Assisted Reproductive Technology

目的:观察2种不同激活方案对辅助生殖技术中未受精的成熟卵母细胞的激活效果及其胚胎发育情况。方法:收集常规IVF及ICSI后48h移植当日仍未受精的成熟卵母细胞,分别采用"Ca2+载体A23187联合6-甲基嘌呤(6-DMAP)"(Ca+6D)和"氯化锶联合6-DMAP"(Sr+6D)2种方案进行激活处理,观察原核形成情况;将激活后的卵母细胞继续体外培养3～5d,观察胚胎发育情况。结果:Ca+6D组的激活率为80.5%(29/36),显著高于Sr+6D组的47.8%(22/46)(P<0.05)。Ca+6D组激活的29个卵母细胞中有15个表现为2PN,占51.7%;Sr+6D组被激活的22个卵母细胞中有10个表现为2PN,占45.4%,组间无统计学差异(P>0.05)。激活的胚胎继续培养,Ca+6D组的卵裂率为48.3%(14/29),2个发育到2～4-细胞,6个发育到5～8-细胞,6个发育到>8-细胞阶段,最终获得4个桑椹胚,1个囊胚;Sr+6D组的卵裂率为31.8%(7/22),2个发育到2～4-细胞,4个发育到5～8-细胞,1个发育到>8-细胞阶段。结论:2种激活方案均能激活辅助生殖技术中未受精卵母细胞,并继续发育形成胚胎。激活方案"Ca2+载体A23187联合6-DMAP"与"SrCl2联合6-DMAP"相比,激活率及激活胚胎的发育结局,前者优于后者,可能是更适合辅助生殖技术中未受精卵母细胞的激活方案。

Objective：To investigate the effect of two different methods on activation of human unfertilized oocyte after ICSI or IVF.Methods：Unfertilized oocytes on the day of embryo transfer at 48 h after ICSI or IVF were allocated to two groups randomly.In group Ca＋6D,oocytes were exposed to 5μmol/L calcium ionophore A23187 for 5 min in dark,and subsequently to 2 mmol/L 6-DMAP for 3 h.In group Sr＋6D,oocytes were exposed to 10 mmol/L strontium chloride（SrCl2）for 20 min,and subsequently to 2.5 mmol/L 6-DMAP for 1.5 h.The oocytes were checked for pronucleus formation at 18-20 h after activation.The activated oocytes were cultured in vitro for 3-5 d.Results：The activation rate of group Ca＋6D（80.5%）was higher than that of group Sr＋6D（47.8%）,P〈0.05.There was no significant difference in the formation rate of 2PN（51.7%in group Ca＋6D vs 45.4%in group Sr＋6D）and the cleavage rate（48.3%in group Ca＋6D vs 31.8%in group Sr＋6D）between the two groups（P〈0.05）.Among the 14 cleaved oocytes in group Ca＋6D,2 developed to 2-4-cell stage,6 developed to 5-8-cell stage,6 developed to 〉8-cell stage,4 morulae and 1 blastocyst were obtained.Among the 7 cleaved oocyteds in group Sr＋6D,2 devel-oped to 2-4-cell stage,4 developed to 5-8-cell stage,1 developed to〉8-cell stage.Conclusion：Unfertilized oocytes after IVF or ICSI 48 h could be artificially activated and developed to embryos.The treatment of the combination of calcium ionophore A23187 with 6-DMAP was better than that of the combination of SrCl2 with 6-DMAP in respects of activation rate and the development of the activated embryos.