Ca2+载体A23187对受精失败人类成熟卵母细胞的补救激活作用

目的:观察Ca²⁺载体A23187对受精失败人类成熟卵母细胞的补救激活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24h仍无受精征象的成熟卵母细胞作为研究对象,5μmol/LCa²⁺载体A23187中处理5min,观察第二极体排出及原核形成情况,激活后卵母细胞在体外继续培养2d。结果:IVF组及ICSI组卵母细胞激活率分别为64.9%(24/37)和73.2%(30/41)。ICSI组受精失败卵母细胞激活后主要表现为二极体二原核(2PB+2PN)(80%,24/30),而IVF组仅有20%的被激活卵母细胞表现为2PB+2PN,两组间差异具极显著(P＜0.01)。31个2PN卵母细胞继续培养,25个发生分裂,11个发育到2-4细胞,8个发育到4-8细胞,6个发育到8-细胞以上阶段。结论:Ca²⁺载体A23187能够有效地激活ICSI后受精失败卵母细胞恢复受精并继续发育成胚胎。

Rescue activation of calcium ionophore A23187 on unfertilized human oocytes after conventional in-vitro fertilization and intracytoplasmic sperm injection

AIM:To explore the activation effect of calcium ionophore A23187 on unfertilized human mature oocytes after conventional in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI).METHODS:Thirty-seven unfertilized mature oocytes from IVF and 41 after ICSI were included in our experiment. They were incubated in 5 μmol/L calcium ionophore A23187 for 5 minutes. Second polar body extrusion and pronuclear formation were recorded 12-16 hours later. The activated oocytes were cultured for another 2 days in vitro.RESULTS:Activation rate of unfertilized oocytes from conventional IVF and ICSI were 64.9%(24/37)and 73.2%(30/41), respectively. Among 41 unfertilized oocytes after ICSI treated with A23187, 30 were activated and 24 had 2 polar body (PB) and 2 pronuclear (PN). But for the unfertilized oocytes from conventional IVF only 20% activated oocytes had 2 PN and 2 PB. The percentage difference of oocytes containing 2 PB and 2 PN between the two groups was significant (P＜0.01). When all 31 activated oocytes containing 2 pronuclei were cultured in IVF-20 for additional 48 hours, 25 cleaved. Among them 11 developed to 2-4 cell stage, 8 to 4-8 cell stage and 6 to 8-cell stage.CONCLUSION:Calcium ionophore A23187 was effective in activating unfertilized human oocytes after IVF or ICSI. The activated oocytes had further developmental potential.