Kierszenbaum A L, DePhilip R M, Spruill W A, Takenaka I
Exp Cell Res. 1983 May;145(2):293-304. doi: 10.1016/0014-4827(83)90008-3.
A method for the isolation and culture of seminal vesicle epithelial cells obtained from control and androgen-primed sexually-immature, uncastrated rats is described. This method allows the establishment of monolayer cultures from aggregates of seminal vesicle epithelial cells isolated after trypsin and collagenase digestion. Phase contrast and transmission electron microscopic methods demonstrate that cell aggregates, after attaching to the substrate, establish within 48 h a colony-like, epithelial-like growth pattern. Immunofluorescent localization studies of SVS IV, an androgen-dependent secretory protein purified from rat seminal vesicle secretion, show that cultured seminal vesicle epithelial cells are immunoreactive. An electrophoretic analysis of [35S]methionine-labeled secretory proteins immunoprecipitated with rabbit anti-SVS IV serum demonstrate that, whereas SVS IV is newly-synthesized and accumulated in the medium of cultured seminal vesicle cells established from androgen primed rats, cultured cells from control rats appear to synthesize and accumulate SVS IV in a precursor form. Results of this work show that seminal vesicle epithelial cells in culture not only retain several structural features representative of the tissue but also serve as a potential system for the study of androgen action.
本文描述了一种从对照及经雄激素预处理的性未成熟、未阉割大鼠中分离和培养精囊上皮细胞的方法。该方法可从经胰蛋白酶和胶原酶消化后分离得到的精囊上皮细胞聚集体建立单层培养。相差显微镜和透射电子显微镜方法表明,细胞聚集体附着于底物后,在48小时内形成集落样、上皮样生长模式。对从大鼠精囊分泌物中纯化的雄激素依赖性分泌蛋白SVS IV进行免疫荧光定位研究表明,培养的精囊上皮细胞具有免疫反应性。用兔抗SVS IV血清免疫沉淀的[35S]甲硫氨酸标记分泌蛋白的电泳分析表明,虽然SVS IV在从经雄激素预处理的大鼠建立的培养精囊细胞培养基中是新合成并积累的,但对照大鼠的培养细胞似乎以前体形式合成并积累SVS IV。这项工作的结果表明,培养的精囊上皮细胞不仅保留了该组织的几个代表性结构特征,而且还可作为研究雄激素作用的潜在系统。