山羊卵巢无腔卵泡卵母细胞的体外生长

Growth of Hircine Ovarian Nonantral Follicle Oocytes Xin vitro

山羊卵巢无腔卵泡卵母细胞在以ＤＭＥＭ为基础，添加ＨＥＰＥＳ（２０ｍｍｏｌ／Ｌ）、ＦＣＳ（１０％）、ＦＳＨ（４０ｍｇ／Ｌ）、次黄嘌呤（２ｍｍｏｌ／Ｌ）、异双丁酰环腺苷酸（２ｍｍｏｌ／Ｌ）、ＩＧＦ－Ｉ（５０μｇ／Ｌ）、氢化可的松（４０μｇ／Ｌ）和ＩＴＳ（５０μｇ／Ｌ）的培养液中得以存活并生长。在二维培养体系中，卵泡在体外的生长模式和体内有很大差别。最显著的特征是卵泡不像在体内那样保持完整的立体结构一直到结束。绝大部分卵泡不同程度地发生基膜溶解和破裂，颗粒细胞向四周扩展并贴壁，形成单层。由于卵泡原有三维立体结构的破坏，易于导致卵母细胞的迁移。在生长方式上以数个卵泡聚集生长对其卵母细胞的生长似乎较为有利。卵泡卵母细胞在体外培养９ｄ，其直径可达１５０μｍ以上，达到了成熟时体积，存活率为５３％。实验证明，山羊无腔卵泡卵母细胞在合适的培养体系中，能在体外存活并生长，并能发育到成熟时的大小。

The hircine nonantral follicle oocytes were proved in the present experiment to survive and grow in DMEM supplemented with HEPES (20 mmol/L), FCS(10%), FSH(40 mg/L), hypoxanthine (2 mmol/L), dbcAMP(2 mmol/L), IGF1(50 g/L), hydrocortisone(40 g/L) and ITS(50 g/L). In a twodimensional culture system, the in vitro growth pattern of the follicles in a heap seemed to be of benefit to the oocytes. After the follicle oocytes were cultured in vitro for 9 days the oocytes had grown up to be over 150 m in diameter, reaching the maturation size with the survival rate of 53%. The experiment demostrated that the hircine nonantral follicle oocytes may survive and grow in vitro in a suitable culture system up to their maturation size.