中文摘要：

目的通过对单个卵裂球的体外培养发育的观察,评估其细胞增殖、囊胚形成以及细胞发育分化。方法收集临床体外受精周期中第3天新鲜废弃胚胎,分离单个卵裂球后即移至囊胚培养液中培养。观察卵裂球的生长情况,分析体外发育单个卵裂球的增殖情况、囊胚的形成时间以及效率,并对形成的囊胚进行细胞染色计数,总结单个卵裂球体外培养的发育规律;并且通过免疫荧光检测囊胚的多能性标记物Oct3/4的表达,初步探讨单个卵裂球的发育极性。结果收集第3天新鲜废弃胚胎,共分离并进入研究的单个卵裂球596个,卵裂球形成细胞融合在分离后的48h,囊胚形成则在分离后的72 h,而且生成的囊胚体积较小,大部分内细胞团结构不明显,卵裂球增殖率分别34.1%;囊胚形成率为8.6%,而同期收集的废弃胚胎的囊胚形成率为28.7%,组间差异有统计学意义（P〈0.001）;对分离卵裂球培养第4天形成的囊胚进行细胞计数,中位细胞数为24.6;对所形成的囊胚分析Oct3/4的表达发现,只有25.0%囊胚可表达多能性标志物（Oct3/4）,而且囊胚表达多能性标记物的细胞数很少（2-5个）。结论普通胚胎发育规律比较,单个卵裂球的囊胚形成时间延迟1d,囊胚形成率较未分离的废弃胚胎低下;而且单个卵裂球的分离培养可能对细胞的全能性或者极性发育存在负面的影响。

英文摘要：

Objective To investigate the development algorism of single blastomere from discarded embryos. Methods Blastomeres from Day 3 discarded embryos were cultured in vitro. Cells proliferation, blastocysts formation and ICM of blastomeres were observed and recorded. The general development of single blastomere was summarized. Day 4 blastomere - derived blastocysts were stained with Hoechst33342 and the total cell count was determined under fluorescence microscope. The expression of pluripoteney marker Oct3/4 of blastocyst cultured from single blastomere were detected by immunofluorescence. Results A total of 596 blastomeres from discarded embryos were collected. The day of blastomeres separation was defined as Day 1. The development of the majority of the blastomeres followed the biological clock as compaction on Day 3, cavitation on Day 4 and expanded blastocyst on Day 5. The development of blastomere - derived blastocyst was delayed by 1 day when compared to the normal embryonic development. Blastomere - derived blastocysts had small number of TE cells and only a small part of blastocysts had obvious ICM. Only a small part of the blastocysts expressed Oct3/4 and the Oct3/4（ ＋ ） cell count ranged from 2 to 5 in one blastocyst. The Oct3/4（ ＋ ） blastoeyst rate was 25.0%. Conclusion The development of blastomere - derived blastocysts is delayed by 1 day when compared to the normal embryonic development. Culture of isolated blastomere may have negative effect on the pluripotency of the development of blastomeres.