Seto-Young Donna, Leonardi Olga, Park Alice, Holcomb Kevin, Salehi Marzieh, Chang Peter, Yih Melissa, Rosenwaks Zev, Poretsky Leonid
Division of Endocrinology, Department of Medicine, Beth Israel Medical Center and Albert Einstein College of Medicine, NY 10003, USA.
Horm Res. 2005;64(5):238-47. doi: 10.1159/000089349. Epub 2005 Oct 25.
We repeatedly established a nontransformed steroidogenically active human ovarian cell culture derived from oophorectomy specimens. The cells maintained steroidogenic activity for 3-5 passages (6-8 weeks) and responded to stimulation by insulin and gonadotropin. With pregnenolone as substrate, LH stimulated progesterone production up to 124% and FSH up to 121%. Insulin alone stimulated progesterone production up to 135%, in the presence of LH up to 191%, and in the presence of FSH up to 170%. With dehydroisoandrosterone (DHA) as substrate, insulin alone stimulated testosterone production up to 117%, and in the presence of LH (but not FSH) up to 125%. With androstenedione as substrate, insulin alone stimulated estradiol production up to 133%, FSH alone up to 188%, and LH with insulin up to 217%. With progesterone as substrate and in the presence of LH (but not FSH), 17-alpha-hydroxylase activity was stimulated up to 131%. With DHA as substrate and in the presence of LH, 3-beta-hydroxysteroid dehydrogenase (3-beta-HSD) activity was stimulated up to 139%. With androstenedione as substrate, insulin alone stimulated aromatase activity up to 202%, LH up to 208%, and FSH up to 251%. Under the same conditions, in the presence of LH and insulin, aromatase activity was stimulated up to 342%, and in the presence of FSH and insulin, up to 318%. With testosterone as substrate, insulin alone stimulated aromatase activity up to 122%. With testosterone as substrate, in the presence of LH and insulin, aromatase activity was stimulated up to 136%, and in the presence of FSH and insulin, up to 156%. Immunocytochemistry studies directly confirmed presence of aromatase and 3-beta-HSD in these cultured cells. We conclude that a steroidogenically active nontransformed long-term human ovarian cell culture can be repeatedly established from oophorectomy specimens, providing uninterrupted supply of cultured human ovarian cells for a variety of studies of ovarian physiology.
我们多次从卵巢切除标本中建立了一种未转化的具有类固醇生成活性的人卵巢细胞培养物。这些细胞在3 - 5代（6 - 8周）内保持类固醇生成活性，并对胰岛素和促性腺激素的刺激有反应。以孕烯醇酮为底物时，促黄体生成素（LH）可使孕酮生成增加高达124%，促卵泡生成素（FSH）可使孕酮生成增加高达121%。单独使用胰岛素可使孕酮生成增加高达135%，在LH存在时增加高达191%，在FSH存在时增加高达170%。以脱氢表雄酮（DHA）为底物时，单独使用胰岛素可使睾酮生成增加高达117%，在LH（而非FSH）存在时增加高达125%。以雄烯二酮为底物时，单独使用胰岛素可使雌二醇生成增加高达133%；单独使用FSH可使雌二醇生成增加高达188%；LH与胰岛素共同作用时可使雌二醇生成增加高达217%。以孕酮为底物且在LH（而非FSH）存在时，17 - α - 羟化酶活性增加高达131%。以DHA为底物且在LH存在时，3 - β - 羟类固醇脱氢酶（3 - β - HSD）活性增加高达139%。以雄烯二酮为底物时，单独使用胰岛素可使芳香化酶活性增加高达202%，LH可使芳香化酶活性增加高达208%，FSH可使芳香化酶活性增加高达251%。在相同条件下，在LH和胰岛素存在时，芳香化酶活性增加高达342%；在FSH和胰岛素存在时，芳香化酶活性增加高达318%。以睾酮为底物时，单独使用胰岛素可使芳香化酶活性增加高达122%。以睾酮为底物时，在LH和胰岛素存在时，芳香化酶活性增加高达136%；在FSH和胰岛素存在时，芳香化酶活性增加高达156%。免疫细胞化学研究直接证实了这些培养细胞中存在芳香化酶和3 - β - HSD。我们得出结论，可多次从卵巢切除标本中建立具有类固醇生成活性的未转化的长期人卵巢细胞培养物，为各种卵巢生理学研究提供持续的培养人卵巢细胞供应。
