Effect of preantral follicle isolation technique on in-vitro follicular growth, oocyte maturation and embryo development in mice
Effect of preantral follicle isolation technique on in-vitro follicular growth, oocyte maturation and embryo development in mice
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DOI：
10.1093/humrep/17.8.2152
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发表时间：
2002-08-01
影响因子：
6.1
通讯作者：
Englert, Y
中科院分区：
文献类型：
--
作者：
Demeestere, I;Delbaere, A;Englert, Y
背景：在体外培养之前，使用机械和酶促技术分离腔前卵泡已经被描述过。本研究的目的是评估小鼠腔前卵泡的分离程序对其体外发育的影响。方法：采用机械法或酶法分离卵泡，并使用单个非球形培养系统进行培养。对两组的卵泡发育和类固醇生成、卵母细胞体外成熟和胚胎发育进行了评估。研究结果：培养12天后，机械分离的卵泡有较高的存活率，但较低的窦状腔形成率比酶分离的卵泡。与机械分离相比，酶促卵泡分离与更高的睾酮和雌二醇产量相关。酶法分离后，观察到更强的碱性磷酸酶反应，表明酶法分离的卵泡比机械分离的卵泡有更多的卵泡膜细胞。然而，两种分离技术导致相似的卵母细胞成熟和胚胎发育率。结论：酶促卵泡分离不影响卵泡膜细胞的发育。酶促分离后卵泡类固醇生成增强，但卵母细胞的发育能力与机械分离后获得的相当。
BACKGROUND: The use of mechanical and enzymatic techniques to isolate preantral follicles before in-vitro culture has been previously described. The aim of this study was to assess the effect of the isolation procedure of mouse preantral follicles on their subsequent development in vitro. METHODS: Follicles were isolated either mechanically or enzymatically and cultured using an individual non-spherical culture system. Follicular development and steroidogenesis, oocyte in-vitro maturation and embryo development were assessed for both groups. RESULTS: After 12 days of culture, follicles isolated mechanically had a higher survival rate but a lower antral-like cavity formation rate than follicles isolated enzymatically. Enzymatic follicle isolation was associated with a higher production of testosterone and estradiol compared with mechanical isolation. A stronger phosphatase alkaline reaction was observed after enzymatic isolation, suggesting that follicles isolated enzymatically had more theca cells than those isolated mechanically. However, both isolation techniques resulted in similar oocyte maturation and embryo development rates. CONCLUSIONS: Enzymatic follicular isolation did not affect theca cell development. Follicular steroidogenesis was enhanced after enzymatic isolation but the developmental capacity of oocytes was comparable to that obtained after mechanical isolation.