人窦前卵泡的体外发育及其卵母细胞成熟过程的观察

目的探讨在体外培养条件下,人窦前卵泡发育及其卵母细胞成熟的可能性。方法建立适宜的体外培养环境,应用正交设计将不同剂量的人绝经期促性腺激素（ｈＭＧ）、卵泡液、表皮生长因子（ＥＧＦ）组合,作用于来自体外受精－胚胎移植（ＩＶＦ－ＥＴ）抽吸的卵泡液及冲洗液中获得的窦前卵泡。结果窦前卵泡于培养第６～１２天形成窦卵泡,培养２１～２８天时直径≥３００μｍ的卵泡数显著增多（Ｐ＜０．００５）,当以ｈＭＧ１５０ＩＵ／Ｌ、卵泡液４０％、ＥＧＦ６μｇ／Ｌ作用于卵泡时,卵母细胞排出率与再经培养后成熟率均达最高,分别为８３％和３８％（Ｐ＜０．００５）。结论在适宜的培养条件下,来自ＩＶＦ－ＥＴ病人的抽吸卵泡液的窦前卵泡在体外能够发育,且其卵母细胞可达到成熟,是一新的人卵来源。窦前卵泡的体外发育和体内不同

Development of human preantral follicles and oocytes maturation in vitro

OBJECTIVE: To study the development of human preantral follicles and oocytes maturation by in vitro culture. METHODS: Preantral follicles obtained from follicular fluid during egg retrieval in an in vitro fertilization (IVF) program was first incubated in Ham's F10 with 15% fetal cord serum (FCS). After 28 days, the medium was supplemented with different doses of hMG, mature follicular fluid (mFF) and epidermal growth factor (EGF) by orthogonal design. Induction of final maturation was completed in the presence of human menopasual gonadotropin hMG and mFF. RESULTS: Development from preantral to antral follicles was found within 6-12 days of culture. With increasing time, the proportion of follicles with diameters of more than 300 micrograms increased at 21-28 days of culture (P < 0.005). The maximal number of oocytes extrusion and first polar body formation occurred in the presence of 1501U/L hMG, 40% (v/v) mFF and 6 micrograms/L EGF. CONCLUSIONS: Follicular fluid obtained during egg retrieval in an IVF program was found to contain many preantral follicles which could develop into antral follicles with extrusion of oocytes in culture, and the oocytes matured in vitro. However, the course of development of follicles and maturation of oocytes in vitro is quite different from that in vivo.