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目的 :观察钙离子载体 A2 3 1 87( Ca-A2 3 1 87)及 6-甲基嘌呤 ( 6-DMAP)对人类卵母细胞的孤雌激活作用。方法 :收集体外受精周期中不适于进行卵胞浆内单精子注射的生发泡期或第一次减数分裂中期的不成熟卵母细胞 2 44个在体外培养成熟 ,其中 1 55个体外成熟卵母细胞按不同激活方案分组 :5μmol/ L Ca-A2 3 1 87组、1 0 μmol/ L Ca-A2 3 1 87组、对照组、6-DMAP组及 Ca-A2 3 1 87+6-DMAP组。激活处理后 1 2～ 1 8h观察第二极体排出及原核形成情况。结果 :5及 1 0 μmol/ L Ca-A2 3 1 87组卵母细胞激活率分别为 40 .7% ( 1 1 / 2 7)和 3 7.1 % ( 1 3 / 3 5) ,较对照组的 9.5% ( 2 / 2 1 )明显增加 ( P<0 .0 1 ) ;6-DMAP组的激活率 ,1 1 .5% ( 3 / 2 6)较 Ca-A2 3 1 87+6-DMAP组 ,58.7% ( 2 7/ 46)明显下降。 Ca-A2 3 1 87激活后卵母细胞主要表现为一原核二极体 ,而 Ca-A2 3 1 87+6-DMAP激活的卵母细胞以一原核一极体占多数 ( 1 9/ 2 7,70 .4% )。结论 :卵母细胞孤雌激活的发生及原核形成类型与激活方案有关 ,单独 Ca-A2 3 1 87或与蛋白激酶抑制剂 6-DMAP联合应用都能使人类卵母细胞发生孤雌激活 ,二者联合应用更易于诱导卵母细胞发生孤雌激活
Objective: To observe the parthenogenetic activation of human oocytes induced by calcium ionophore A2 3 1 87 (Ca-A2 3 1 87) and 6-methylpurine (6-DMAP). METHODS: 2 44 immature oocytes, which were not suitable for intracytoplasmic sperm injection during in vitro fertilization, or mid-meiotic metaphase, were collected and cultured, of which 1 55 in vitro mature eggs The mother cells were divided into groups according to different activation schemes: 5μmol / L Ca-A2 3 1 87 group, 10 μmol / L Ca-A2 3 1 87 group, control group, 6-DMAP group and Ca-A2 3 1 87 + 6-DMAP group. Activation after treatment 1 2 ~ 1 8h observe the second polar body excretion and prokaryotic formation. Results: The activation rates of 5 and 10 μmol / L Ca-A2 3 1 87 group were 40.7% (1 1/27) and 31.1% (13/35), respectively. Compared with the control group 9.5% (2/2 1) increased significantly (P <0.01). The activation rate of 6-DMAP group was higher than that of Ca-A2 3 1 87 + 6-DMAP group , 58.7% (27/46) decreased significantly. The oocyte activated by Ca-A2 3 1 87 mainly showed a prokaryotic diode, while the oocyte activated by Ca-A2 3 1 87 + 6-DMAP was mainly composed of a prokaryotic polar body (1 9/2 7 , 70.4%). CONCLUSIONS: The parthenogenetic activation of oocytes and the type of prokaryotic activation are related to the activation scheme. Ca-A2 3 1 87 alone or combined with protein kinase inhibitor 6-DMAP can activate parthenogenetic oocytes in human oocytes. Combined application of more easy to induce oocytes in parthenogenetic activation