应用钙离子载体A23187及6-甲基嘌呤对人卵母细胞进行孤雌激活

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

Parthenogenetic activation of calcium ionophore A23187 and 6-dimethylaminopurine on human oocytes

Objective: To investigate the parthenogenetic activation effects of calcium ionophore A23187 (Ca-A23187) and 6-dimethylaminopurine (6-DMAP) on human oocytes. Methods: Immature oocytes at germinal vesicle stage or metaphase of first meiosis from intracytoplasmic sperm injection cycles were harvested and cultured in vitro. One hundred and fifty-five mature oocytes from in vitro maturation were divided into 5groups: 5 μmol/L Ca-A23187 group, 10 μmol/L Ca-A23187 group, DMSO control group, 2.5 mmol/L 6-DMAP group and Ca-A23187+6-DMAP group. Pronucleus formation and second polar body extrusion were recorded 12~18 hours after activation. Results: The activation rates in 5 and 10 μmol/L Ca-A23187 groups were 40.7% (11/27) and 37.1%(13/35) respectively. They were much higher than that in DMSO control group (9.5%,2/21) (P<0.01). Of 26 oocytes treated with 6-DMAP, only 3 showed signs of activation, the activation rate (11.5%,3/26) was significantly lower than that in Ca-A23187+6-DMAP group (58.7%,27/46). Most of the activated oocytes had 1 pronucleus (PN) and 2 polar bodies (PB) in 5 μmol/L Ca-A23187 group and 10 μmol/L Ca-A23187 group. On the other hand 70.4%(19/27) of the activated oocytes in Ca-A23187+6-DMAP group contained 1 PB and 1 PN. Conclusions: Pronucleus formation and activation status were associated with activation protocols. Fresh human oocytes can be activated by Ca-A23187 or the combination of Ca-A23187 and 6-DMAP. Human oocytes were prone to be activated under the combination of Ca-A23187 and 6-DMAP.