显微受精废弃的人类卵母细胞体外成熟及孤雌激活

In vitro Maturation of Human Oocytes Discarded after Intracytoplasmic Sperm Injection Treatment and Parthenogenetic Activation

以卵胞浆单精注射(intracytoplasmic sperm injection,ICSI)后废弃的未成熟人类卵母细胞(生发泡期卵母细胞(the germinal vesicle,GV)和第一次减数分裂中期卵母细胞(the metaphase,MI))为材料,使用卵母细胞体外成熟培养液培养未成熟的卵母细胞,分别在人类绒毛膜促性腺激素(human chorionic gonadotrophin,hCG)注射后45、60、84 h观察卵母细胞成熟情况.分别使用钙离子载体(calcium ionophore,CI)A23187联合6-二甲基氨基嘌呤(6-DMAP)法或精子提取物卵胞质内注射(sperm extracts intracytoplasmic injection,SEII)法两种不同的激活方法对体外成熟MII的卵母细胞进行孤雌激活,评价其体外发育潜能.MI卵子体外成熟率要显著高于GV(75.2%vs 30.6%)(P<0.01).与CI/6-DMAP法相比使用SEII/6-DMAP法在激活率(87.5%vs 70.2%)上要明显高于CI/6-DMAP法(P<0.05),但在卵裂率(65.7%vs 72.5%)和桑囊率(0%vs 5.0%)上SEII/6-DMAP法要低于CI/6-DMAP法.注射hCG 45 h组的卵母细胞激活率(91.3%vs 57.9%)、卵裂率(85.7%vs 57.9%)及桑囊率(9.5%vs 0%)均显著高于注射hCG 60 h组(P<0.01).56.8%(117/206)的ICSI废弃的未成熟卵母细胞可以在体外发育成熟,激活后具有一定的发育潜能,卵龄对卵母细胞的质量和发育能力影响较大.

In vitro maturation of human immature oocytes（GV and MI） discarded after intracytoplasmic sperm injection（ICSI） treatment were observed at 45,60,84 h after human chorionic gonadotrophin（hCG） injection.MII oocytes were activated with two activated methods： calcium ionophore（CI） A23187 and 6-DMAP method and sperm extracts intracytoplasmic injection（SEII） method,and the developmental potential was also evaluated.The rate of in vitro maturation for MI oocytes was significantly higher than that of GV oocytes（75.2% vs 30.6%）（P0.01）.The activation rate of CI/6-DMAP group was higher than that of SEII/6-DMAP group（87.5% vs 70.2%）（P0.05）,but the cleavage rate and the morula/blastocyst devel-opment rate were lower.The activation rate,cleavage rate and the morula/blastocyst development rate of parthenogenones in injection hCG 45 h group were significantly higher than that in injection hCG 60 h group.56.8%（117/206） immature oocytes discarded after ICSI treatment can develop to MII which have the potentiality to develop to blastocyst after parthenogenetic activation.However,the age of oocytes greatly impact on the quality and developmental competence of oocytes.