受精液平衡方式对新建胚胎实验室鼠胚实验的影响

Effect of balance mode of fertilization culture medium on mouse embryo experiment in new embryo laboratory

目的探讨授精当日配皿与前日配皿对昆明小白鼠卵母细胞体外受精及胚胎发育的影响,为人类辅助生殖技术(ART)胚胎实验室选择最佳配皿方案提供参考。方法选择SPF级昆明小白鼠进行实验,对同一批次雌鼠统一超促排卵后按照随机原则分为当日配皿组(当日配制授精皿)与前日配皿组(前日配制授精皿)进行鼠胚实验。授精前1日配制含10%SPS的Quinn’s 1020液,置于37℃、5%CO 2、5%O 2的饱和湿度培养箱中过夜平衡后当日配皿为当日配皿组;于授精前1日配制含10%SPS的Quinn’s 1020液,并直接配皿后置于37℃、5%CO 2、5%O 2的饱和湿度培养箱过夜平衡为前日配皿组。观察并比较两组不同受精液平衡方式中小鼠卵母细胞的受精及胚胎发育情况。结果当日配皿组完成124个小鼠促排卵周期,总获卵4731枚,受精率为68.2%,卵裂率97.7%,囊胚形成率76.7%;前日配皿组完成80例促排卵周期,总获卵2683枚,受精率为84.5%,卵裂率99.6%,囊胚形成率88.8%。前日配皿组的受精率、卵裂率和囊胚形成率均显著高于当日配皿组(P<0.01)。结论受精培养液的平衡方式对昆明小白鼠卵母细胞的受精及胚胎发育可产生显著影响,前日配皿较当日配皿更有利于小鼠卵母细胞的受精及后期的胚胎发育。

Objective:To explore the effects of balance mode of fertilization culture medium on mouse embryo experiment in new embryo laboratory.Methods:Kunming mice of SPF grade were selected for the experiment.The female mice of the same batch were randomly divided into two groups after ovarian hyperstimulation:same day-made group(insemination dishes made on the same day)and previous day-made group(insemination dishes made on the previous day).The Quinn’1020 solution containing 10%SPS were prepared one day before insemination,and the insemination dishes were prepared on the same day after overnight balancing in a saturated humidity incubator with 37℃,5%CO 2 and 5%O 2 in same day-made group.The Quinn’1020 solution containing 10%SPS were prepared and directly prepared the insemination dishes on the day before insemination,then placed the dishes in a saturated humidity incubator at 37℃,5%CO 2 and 5%O 2 for overnight balance in previous day-made group.The fertilization and embryonic development of mouse oocyte in different fertilizer balance modes were observed and compared between the two groups.Results:There were 124 ovulation induction cycle,with a total of 4731 oocytes,fertilization rate 68.2%,cleavage rate 97.7%and blastocyst formation rate 76.7%in the same day-made group,and 80 ovulation induction cycle,with a total of 2683 oocytes,fertilization rate 84.5%,cleavage rate 99.6%and blastocyst formation rate 88.8%in previous day-made group.The fertilization rate,cleavage rate and blastocyst formation rate of the previous day-made group were significantly higher than those of the same day-made group(P<0.01).Conclusions:The balance mode of fertilization medium has a significant effect on the fertilization and embryonic development of Kunming mouse oocytes.The insemination dishes made on previous day are more beneficial to fertilization of mouse oocyte and later embryonic development than that the insemination dishes made on the same day.