人卵母细胞的电脉冲辅助激活

Assisted Activation of Electrical Stimulation on Human Mature Oocytes

[目的]探讨电脉冲刺激进行人卵母细胞的人工辅助激活的可行性。[方法]不同卵龄和来源的人卵母细胞分为4组,49个新鲜成熟的卵母细胞作为A组,42个常规体外受精(IVF)中未受精的成熟卵母细胞为B组,C组为卵浆内单精子注射(ICSI)后未受精卵母细胞57个,D组为体外培养48h的ICSI后未受精卵母细胞37个,所有卵母细胞以直流电脉冲刺激2次,观察电脉冲刺激后卵母细胞激活和胚胎发育情况。[结果]A组卵母细胞激活率为43％(21/49),与B、C、D3组相比偏低,差异具有统计学意义(P均<0.05),A组中激活后卵母细胞主要表现为一原核二极体(IPN+2PB),而C组和D组则以二原核二极体(2PN+2PB)占多数。A、B、C组激活后卵母细胞分裂率分别为81％(17/21),83％(24/29)和88％(38/43),较D组(19％,5/26)显著增加(P<0.01),在电脉冲刺激后第3天,A、B、C组分别有18％(3/17),29％(7/24),31％(11/35)的胚胎发育到8-细胞阶段。[结论]不同来源和卵龄的人卵母细胞在电脉冲刺激后表现为不同的激活类型和胚胎发育情况,直流电脉冲作为一种有效的卵母细胞人工激活方法可以应用于辅助受精。

[Objective] To observe activation effects of electrical stimulation on human mature oocytes. [ Methods ] Oocytes with different ages were divided into four groups. Forty-nine fresh oocytes after vitro maturation were obtained in group A. In group B unfertilized culture-aged oocytes after conventional in vitro fertilization were recruited ( n - 42). Fifty-seven unfertilized oocytes after ICSI were included in group C. And in group D unfertilized oocytes after 48 hours culture in. vitro were collected ( n = 37). All oocytes were stimulated two times with direct current pulse. Oocyte activation and embryo development were observed. [Results] Activation rate in group A was 42. 9% (21/49), significantly lower than that in group B, group C, and group D (69.0% -75.4%, P <0. 05). Two thirds of activated oocytes ( n = 21) in group A displayed one pronuclear (PN) and two polar bodies (PB) (1PN + 2PB) . In contrast in group C and group D oocytes with two pro nuclei and two polar bodies (2PN + 2PB) were found as the predominant pattern. The cleavage rates in group A, group B, and group C were 81.0% (17/21), 82.8% (24/29) and 88.4% (38/43) respectively, much higher than that in group D (19. 2%, 5/26) (P <0.01). On the third day after electrical stimulation, 17.6% (3/ 17), 29.2% (7/24), 31.4% (11/35) of activated oocytes progressed to 8-cell stage respectively in group A, group B and group C. [Conclusion] Human oocytes at different ages can be effectively activated by electrical stimulation, with different activation patterns and embryo development. As an effec- tive activation method, direct current pulse may be used to assist fertilization .