应用人黄素化颗粒细胞条件培养液行人类胚胎体外培养

目的 研究卵泡壁颗粒细胞条件培养液在人类胚胎体外发育中的作用。方法 将卵泡液中回收的卵泡壁颗粒细胞，分别置于添加0．1％聚乙烯醇(PVA)和添加10％胎牛血清(FCS)的TCM-199培养系统中进行体外培养。结果 在无蛋白培养系统中，颗粒细胞能够缓慢生长，第三～五天达到生长高峰，群体倍增时间发生在培养的第二～三天，培养第一～五天可分泌雌二醇到培养液中(3．22～4．65pmol／ml)。在有蛋白培养系统中，颗粒细胞生长速度明显高于无蛋白的培养系统。使用颗粒细胞条件培养液，获得的囊胚在囊胚腔的大小和细胞数量上，均优于商品化囊胚培养液。结论颗粒细胞条件培养液有利于人类4-细胞以后胚胎的发育。

The culture of human embryos in homologous granulosa cells conditioned medium

Objective To investigate the function of human granulosa cell conditioned medium on human embryos development in vitro. Methods Mural luteinizing granulosa cells were collected from follicular fluid when oocytes were retrieved from IVF patients. Digested by trpsin, human granulosa cells were cultured in TCM-199 with 0.1% PVA or 10% FCS in vitro. Results It showed that, in the present of protein, the granulosa cells grew faster than in protein-free medium. Granulosa cells cultured in protein-free system secreted estradiol into medium. The estradiol content in the medium cultured for 1-5 days was significantly higher than those in control medium, but there was no significance between different culture dates. The number of granulosa cells reached the peak at day 3-5 of culture. The multiple period is day 2-3 of culture. Whereas, day 1 and day 5-6 of culture was the slow growth period. By culture of human 4-cell stage embryos in granulosa cell conditioned medium, the blastocyst rate reached 23.3%, lower than that in commercial blastocyst medium (33.3%), but the expanded blastocyst rate and the blastocyst cell number were bigger in the granulosa cell conditioned medium than in the commercial blastocyst medium. Conclusion Granulosa cell conditioned medium is beneficial for human embryos development at 4-cell stage or later stages.