兔卵母细胞胞质内单精子注射后二次辅助激活对兔卵母细胞发育能力的影响

目的 观察兔卵母细胞胞质内单精子注射(ICSI)后2次辅助激活对兔卵母细胞发育能力的影响.方法 采集兔卵母细胞500枚,玻璃化冷冻后解冻,行ICSI后培养1 h,将存活的156枚卵母细胞分为10634激活1次组(30枚,加入浓度为5 μmoL/L的钙离子载体10634激活5 min)、氯化锶激活1次组(26枚,加入浓度为10 mmol/L的氯化锶激活20 min)、10634激活2次组(33枚,加入浓度为5μmoL/L的钙离子载体10634激活5 min后培养30 min,采用同样方法再次激活)、氯化锶激活2次组(28枚,加入浓度为10 mmol/L的氯化锶激活20 min后培养30 min,采用同样方法再次激活)和对照组(39枚,未加入任何激活剂)共5组,观察各组卵母细胞的受精率、卵裂率和囊胚形成率.结果 各组兔卵母细胞的受精率、卵裂率和囊胚形成率,氯化锶激活1次组分别为54%、27%和8%,均高于10634激活1次组(分别为33%、17%和3%)和对照组(分别为33%、18%和3%),但差异均无统计学意义(P>0.05);10634激活2次组分别为82%、55%和15%,均高于10634激活1次组,差异均有统计学意义(P<0.05);氯化锶激活2次组的受精率(61%)高于氯化锶激活1次组,卵裂率和囊胚形成率(分别为21%和7%)均低于氯化锶激活1次组,但差异均无统计学意义(P>0.05);10634激活2次组的受精率、卵裂率和囊胚形成率均高于氯化锶激活2次组,差异均有统计学意义(P<0.05).结论 冻融的兔卵母细胞ICSI后行2次辅助激活,可能会提高卵母细胞的受精率和胚胎早期的发育能力,但是激活后的卵母细胞分裂速度较快.

Effect of double activation on the development of frozen-thawed oocytes after intracytoplasmic sperm injection

Objective To investigate the influence on developmental potential of frozen-thawed rabbit oocytes with double assisted activation followed by intracytoplasmic sperm injection (ICSI). Methods A total of rabbit oocytes were collected and thawed after vitrification cryopreservation. Among all oocytes were cultured for 1 hour followed by ICSI. 156 Survived oocytes were divided into 5 groups randomly. I0634 single activation: 30 oocytes were added with calcium ionomycin ( I0634 ) at 5 μmol/L for 5 minutes;SrCl2single activation: 26 oocytes were added with strontium chloride at 10 mmol/L for 10 minutes;10634 double activation: 33 oocytes were activated by I0634 twice;SrCl2 double activation: 28 oocytes were activated by strontium chloride twice. Control group: 39 oocytes were not added with any activators. The rate of fertilization, cleavage and blastocysts formation were observed and compared between various groups. Result The rates of fertilization, cleavage and blastocysts formation were in group of SrCl2 single activation were higher than those of I0634 single activation group without statistical difference (54% vs. 33%, 27% vs. 17%, 8% vs. 3%, P <0.05 ). However, those above rates in double activation by I0634 were higher significantly than those of single I0634 activation (82% vs. 33%, 55% vs. 17%, 15% vs. 3%, P < 0.05). The rates of fertilization (61%) was higher and the rate of cleavage (21%) and blastocysts formation (7%) were lower in group of SrCl2 double activation in comparison with group of SrCl2 single activation without reaching statistical difference (P<0.05 ). Notably, the rates of fertilization, cleavage and blastocysts formation in I0634 double activation group were higher than those in group of SrCl2 double activation with statistical difference (82% vs. 61%, 55% vs. 21%, 15% vs. 7%, P<0.05). Conclusion It might enhance the potential of fertilization of oocytes and early embryo development treated by double activation following ICSI, however, those activated oocytes demonstrate rapid cleavage.