辅助生殖技术中未受精卵母细胞经人工激活剂作用的激活效果观察

Effect of artificial activator on activation of human unfertilized oocytes after assisted reproductive technology

目的观察辅助生殖技术中未受精成熟卵母细胞经钙离子载体A23187联合6-甲基嘌呤(6-DMAP)处理后的激活效果,以及激活后的胚胎发育情况。方法收集常规体外受精(IVF)及卵浆内单精子注射(ICSI)后48h仍未受精的成熟卵母细胞,经5μmol/L钙离子载体A23187处理5min,然后在2mmol/L6-DMAP中处理3h,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 IVF及ICSI后未受精卵母细胞共36个,激活29个,激活率80.5%,1PN形成率31.0%,2PN形成率51.7%,多原核形成率17.2%;继续培养,卵裂率为48.3%(14/29)。15个2PN中有10个发生卵裂,2个发育至2-4细胞阶段,5个发育至5-8细胞阶段,3个发育至>8细胞阶段,最终获得2个桑葚胚。结论辅助生殖技术中未受精卵母细胞可在体外被钙离子载体A23187联合6-DMAP激活,并继续发育形成胚胎。

Objective：To investigate the effect of calcium ionophore A23187 combined with 6-dimethylam inopurine（6 DMAP） on activation of human unfertilized oocytes after intracytoplasmic sperm injection（ICSI） or in intro fertilization（IVF）.Methods：Unfertilized oocytes on the day of embryo transfer at 48h after ICSI or IVF were exposed to 5μmol /L calcium ionophore A23187 for 5 minutes in dark,and subsequently to 2 mmol/L 6-DMAP for 3h.The oocytes were checked for pronucleus formation at 18-20 hours after activation.Fertilized oocytes were cultured in vitro for 3 ～ 5d,the developmental potential of the activated embryos were observed.Results：29 out of 36 unfertilized oocytes were activated,and 51.7% of the activated oocytes show 2PN.Among the 10 cleaved oocyteds,2 developed to 2-4 cell stage,5 developed to 5-8 cell stage,2 developed to morular stage.Conclusion：The treatment of calcium ionophore A23187 combined with 6-DMAP could effectively activate unfertilized oocytes after IVF or ICSI 48h,and the normal fertilized oocytes could develop to embryos.