Lu Qun, Chen Zi-jiang, Gao Xuan, Ma Shui-ying, Li Mei, Hu Jing-mei, Li Yuan
Reproductive Medicine Center, Shandong Provincial Hospital, Shandong University, Jinan 250021, China.
Zhonghua Fu Chan Ke Za Zhi. 2006 Mar;41(3):182-5.
To investigate the effect of assisted oocyte activation with calcium ionophore A23187 and puromycin on human oocytes that fail to fertilize after intracytoplasmic sperm injection (ICSI).
All 113 discarded oocytes that showed no evidence of fertilization at 16 - 18 hours after in vitro maturation (IVM)-ICSI cycles and conventional ICSI were assigned to four groups according to the time after ICSI: IVM-ICSI 22-hour group (n = 33), IVM-ICSI 44-hour group (n = 18), ICSI 44-hour group (n = 37) and ICSI 68-hour group (n = 25). All unfertilized oocytes were exposed to calcium ionophore A23187 (5 micromol/L) for 5 minutes and subsequently incubated with puromycin (10 microg/ml) for 4 hours. After incubation, the oocytes were cultured in vitro for 3 - 5 days. The activation rate, proportion of oocytes that showed pronucleus formation and cleavage rate were calculated after activation. Sex chromosomal analysis was performed by dual color fluorescence in situ hybridization (FISH) on the embryos that displayed two pronuclei and a second polar body.
The combination of calcium ionophore A23187 with puromycin could activate the unfertilized oocytes 22 - 68 hours after ICSI. Best results were achieved in IVM-ICSI 22-hour group, which elicited 88% (29/33) of activation rate, 62% (18/29) of cleavage rate and 28% (5/18) of 4-cell embryos. One embryo in this group developed to the morular stage. The activation rate and developmental potential of the activated embryos in IVM-ICSI 44-hour group, ICSI 44-hour group and ICSI 68-hour group decreased. FISH analysis showed 4 embryos with XX and 9 embryos with XY in 16 embryos.
The combination of calcium ionophore A23187 with puromycin could effectively activate unfertilized oocytes 22 - 68 hours after ICSI. The cultured time of unfertilized oocytes after ICSI affects activation efficiency and developmental potential of the activated embryos. The activated zygotes that display two pronuclei and a second polar body can develop normally.
探讨钙离子载体A23187和嘌呤霉素辅助卵母细胞激活对卵胞浆内单精子注射（ICSI）后未受精的人卵母细胞的影响。
将113枚在体外成熟（IVM）-ICSI周期及传统ICSI后16 - 18小时未显示受精迹象的废弃卵母细胞，根据ICSI后的时间分为四组：IVM-ICSI 22小时组（n = 33）、IVM-ICSI 44小时组（n = 18）、ICSI 44小时组（n = 37）和ICSI 68小时组（n = 25）。所有未受精卵母细胞均暴露于钙离子载体A23187（5微摩尔/升）5分钟，随后与嘌呤霉素（10微克/毫升）孵育4小时。孵育后，将卵母细胞体外培养3 - 5天。激活后计算激活率、显示原核形成的卵母细胞比例和卵裂率。对显示两个原核和第二极体的胚胎进行双色荧光原位杂交（FISH）进行性染色体分析。
钙离子载体A23187与嘌呤霉素联合应用可激活ICSI后22 - 68小时未受精的卵母细胞。IVM-ICSI 22小时组效果最佳，激活率为88%（29/33），卵裂率为62%（18/29），4细胞胚胎率为28%（5/18）。该组有1枚胚胎发育至桑葚胚阶段。IVM-ICSI 44小时组、ICSI 44小时组和ICSI 68小时组激活胚胎的激活率和发育潜能降低。FISH分析显示，16枚胚胎中4枚为XX，9枚为XY。
钙离子载体A23187与嘌呤霉素联合应用可有效激活ICSI后22 - 68小时未受精的卵母细胞。ICSI后未受精卵母细胞的培养时间影响激活胚胎的激活效率和发育潜能。显示两个原核和第二极体的激活合子可正常发育。
