Katayama E
Nihon Sanka Fujinka Gakkai Zasshi. 1984 Jun;36(6):927-36.
The aim of this study was to establish a monolayer culture system for human ovarian thecal cells and to investigate their morphological and functional characteristics. Theca layers were isolated and digested with collagenase-hyaluronidase solution, and dispersed thecal cells were cultured for 10 days in plastic dishes. Histological examination indicated that there were no contaminating granulosa cells in isolated theca layers. Various histochemical studies revealed abundant lipid droplets and 3 beta-hydroxysteroid dehydrogenase activity in cultured cells. The major steroids secreted were delta 4-androstenedione (delta 4) and progesterone(P). delta 4 secretion was very high during the first 2 days (31.6 +/- 1.9 ng/1 X 10(5) cells/2 days) and declined thereafter. P was secreted in moderate amounts throughout the 10 day culture period (9.0-21.3 ng/1 X 10(5) cells/2 days), while estradiol secretion was very low. Subsequently, the responsiveness of cultured thecal cells to gonadotropins and dibutyryl cyclic AMP (Bu2cAMP) was investigated. LH/HCG and Bu2cAMP stimulated delta 4 and P secretion in a dose-related manner. The maximal effective doses of LH and HCG were both 10 ng/ml, and that of Bu2cAMP was 10(-3)M. In conclusion, it was evident that these monolayer-cultured human thecal cells could maintain their morpho-functional characteristics during culture. Therefore, this culture system will provide an excellent model for further studies on the functional properties of thecal cells.
本研究的目的是建立人卵巢膜细胞的单层培养系统，并研究其形态和功能特征。分离卵泡膜层并用胶原酶-透明质酸酶溶液消化，将分散的膜细胞在塑料培养皿中培养10天。组织学检查表明，分离的卵泡膜层中没有污染的颗粒细胞。各种组织化学研究显示，培养细胞中有丰富的脂滴和3β-羟类固醇脱氢酶活性。分泌的主要类固醇为δ4-雄烯二酮（δ4）和孕酮（P）。δ4分泌在最初2天非常高（31.6±1.9 ng/1×10(5)细胞/2天），此后下降。在整个10天的培养期内，P以中等量分泌（9.0 - 21.3 ng/1×10(5)细胞/2天），而雌二醇分泌非常低。随后，研究了培养的膜细胞对促性腺激素和二丁酰环磷酸腺苷（Bu2cAMP）的反应性。LH/HCG和Bu2cAMP以剂量相关的方式刺激δ4和P的分泌。LH和HCG的最大有效剂量均为10 ng/ml，Bu2cAMP的最大有效剂量为10(-3)M。总之，很明显这些单层培养的人膜细胞在培养过程中可以保持其形态功能特征。因此，这种培养系统将为进一步研究膜细胞的功能特性提供一个极好的模型。
