Ikeda H
Department of Biochemistry, Kitasato University School of Medicine, Kanagawa, Japan.
In Vitro Cell Dev Biol. 1990 Feb;26(2):193-200. doi: 10.1007/BF02624112.
Thecal cells isolated from bovine ovarian follicles were cultured with a serum-free basal medium or a serum-free complete medium in the presence or absence of collagen gel matrix, and their cellular proliferation and steroidogenesis were compared with those of cells cultured with a serum-containing medium. The cells cultured with the serum-free basal medium produced larger amounts of progesterone, androstenedione, and estradiol than the cells cultured with the serum-containing medium, but no appreciable cell proliferation was observed in the serum-free medium. Response of thecal cells to 8 bromo-cAMP, a steroidogenic agent, varied according to the type of steroid production examined and the type of culture medium used. In a cultivation period of 4 d, progesterone production was stimulated about five-fold by 8 bromo-cAMP in the serum-free complete medium on collagen gel matrix and in the serum-free basal medium without collagen matrix, whereas androstenedione production was stimulated about three- to fourfold in the serum-free complete medium on collagen gel matrix and in the serum-free basal medium with or without collagen matrix. Estradiol production, however, was significantly suppressed by 8 bromo-cAMP in the serum-free complete medium on collagen gel matrix and also in the serum-containing medium. Thus, among the conditions examined, the most suitable primary culture media for steroidogenesis of thecal cells were the serum-free media, especially serum-free complete medium on collagen gel matrix.
从牛卵巢卵泡中分离出的膜细胞，在有无胶原凝胶基质的情况下，分别用无血清基础培养基或无血清完全培养基进行培养，并将其细胞增殖和类固醇生成情况与用含血清培养基培养的细胞进行比较。与用含血清培养基培养的细胞相比，用无血清基础培养基培养的细胞产生了更多的孕酮、雄烯二酮和雌二醇，但在无血清培养基中未观察到明显的细胞增殖。膜细胞对类固醇生成剂8-溴环磷酸腺苷（8-bromo-cAMP）的反应，根据所检测的类固醇生成类型和所用培养基类型而有所不同。在4天的培养期内，在胶原凝胶基质上的无血清完全培养基和无胶原基质的无血清基础培养基中，8-溴环磷酸腺苷使孕酮生成增加约5倍，而在胶原凝胶基质上的无血清完全培养基以及有或无胶原基质的无血清基础培养基中，雄烯二酮生成增加约3至4倍。然而，在胶原凝胶基质上的无血清完全培养基以及含血清培养基中，8-溴环磷酸腺苷显著抑制了雌二醇的生成。因此，在所检测的条件中，用于膜细胞类固醇生成的最适宜的原代培养基是无血清培养基，尤其是胶原凝胶基质上的无血清完全培养基。
