Yoshimura Y, Hosoi Y, Iritani A, Nakamura Y, Atlas S J, Wallach E E
Department of Obstetrics and Gynecology, Fujita-Gakuen Health University School of Medicine, Aichi, Japan.
Biol Reprod. 1989 Jul;41(1):26-33. doi: 10.1095/biolreprod41.1.26.
The present study was undertaken to define hormonal conditions for in vitro maturation that support subsequent fertilization and embryonic development. Follicular oocytes were recovered from nonstimulated rabbit ovaries and cultured for 12 h in Brackett's medium supplemented with or without hormones. Matured oocytes were inseminated in vitro and transferred 12 h later to Ham's F-10 medium supplemented with 20% fetal calf serum. The initial cleavage frequency of matured oocytes in Brackett's medium was comparable to the frequency of development for in vitro-matured oocytes under various hormonal conditions. However, the addition of estradiol (E2, 1 microgram/ml) to incubation medium containing luteinizing hormone (LH) and follicle-stimulating hormone (FSH) increased significantly (p less than 0.001) the percentage of embryos achieving morula or blastocyst formation (16/98, 16.3%), as compared to the mature oocytes in medium containing LH, LH plus FSH, or no hormone. The addition of prolactin (PRL) to the maturation medium increased the percentage of development to organized embryos in a dose-dependent manner. In vitro-matured oocytes in medium containing LH, FSH, and PRL exhibited a significantly (p less than 0.001) lower incidence of developmental competence (5/95, 5.3%) than oocytes matured in the presence of E2 in conjunction with pituitary hormones (43/89, 48.3%). These results demonstrate that hormonal composition in the environment of the oocyte is critical for acquisition of developmental capacity. PRL as well as E2 appears to be an important constituent in the process of oocyte maturation, promoting preimplantation embryonic development.
本研究旨在确定支持后续受精和胚胎发育的体外成熟激素条件。从未受刺激的兔卵巢中回收卵泡卵母细胞,并在添加或不添加激素的布雷克特培养基中培养12小时。成熟的卵母细胞进行体外受精,12小时后转移至添加20%胎牛血清的哈姆氏F-10培养基中。布雷克特培养基中成熟卵母细胞的初始分裂频率与各种激素条件下体外成熟卵母细胞的发育频率相当。然而,与含促黄体生成素(LH)、LH加促卵泡激素(FSH)或不含激素的培养基中的成熟卵母细胞相比,在含LH和FSH的孵育培养基中添加雌二醇(E2,1微克/毫升)显著增加(p<0.001)了达到桑椹胚或囊胚形成的胚胎百分比(16/98,16.3%)。向成熟培养基中添加催乳素(PRL)以剂量依赖方式增加了发育成有组织胚胎的百分比。与在E2与垂体激素共同存在下成熟的卵母细胞(43/89,48.3%)相比,含LH、FSH和PRL的培养基中体外成熟的卵母细胞的发育能力发生率显著降低(p<0.001)(5/95,5.3%)。这些结果表明,卵母细胞环境中的激素组成对于发育能力的获得至关重要。PRL以及E2似乎是卵母细胞成熟过程中的重要组成部分,促进着床前胚胎发育。