成熟培养时间对家猫卵母细胞孤雌激活胚发育的影响

为筛选出最佳体外成熟时间,以乙醇联合6-DMAP激活验证成熟时间对家猫卵母细胞成熟的影响.将家猫卵母细胞分别体外成熟28h,32h和36h后,采用10%无水乙醇激活3min,6-DMAP培养4h,再移入胚胎体外培养液中培养;第二天观察卵裂率,第六天结束培养并用H33342染色,观察不同成熟时间的激活胚胎发育情况.结果表明:成熟32h组的卵母细胞激活率最高,显著高于28h组(60.42%vs 43.16%,0.010.05);经孤雌激活的卵母细胞没有发育至囊胚的,32h组桑椹胚率最高达到15.63%,与其他两组差异不具有统计学意义(7.36%和10.78%,p>0.05).结论:家猫卵母细胞体外成熟培养32h,成熟效果最好.

EvaluationoftheEffectofMaturationDuration onOocyteDevelopmentPotentialityofDomestic CatUsingArtificialStimulationwithEthanoland6-DMAP

The aim of this paper was to investigate the effect of parthenogenetic activation duration on feline oocyte development capability and to select the best in vitro maturation duration.Feline oocytes were treated for 28 h,32 h and 36 h,respectively,in medium 199 added with FSH and LH for in vitro maturation(IVM).Next,the in vitro maturated oocytes were subjected to artificial activation in 10% ethanol and 6-DMAP.Then,the activated oocytes were cultured for 6 days.The rate of activated oocytes was recorded on Day 2 and the oocytes were stained with Hoechest 33342 on Day 6 to observe their development.The rate of activated oocytes of the 32 h group was significantly higher than that of the 28 h group(60.42% vs 43.16%,p<0.05),but there was no significant difference between the 32 h group and the 36 h group(60.42% vs 50.00%,p>0.05).No oocyte developed to blastula after activation.The rate of morula in the 32 h group was the highest(15.63%),which was,however,not significantly different(p>0.05) from that of the 28 h group(7.36%) or the 36 h group(10.78%).In conclusion,the optimal maturation duration of feline oocytes was 32 h.