Taketa S, Nishi N, Takasuga H, Okutani T, Takenaka I, Wada F
Department of Urology, Kagawa Medical School, Japan.
Prostate. 1990;17(3):207-18. doi: 10.1002/pros.2990170304.
Different procedures of enzymatic digestion of rat prostatic tissue and unique sets of mitogenic factors made it possible to culture practically pure populations of epithelial and stromal cells without previous separation of the two types of cells. Keratin-positive epithelial cells dissociated by trypsin and collagenase from adult rat ventral prostate proliferated in medium WAJC 404 supplemented with epidermal growth factor, insulin, cholera toxin, and bovine pituitary extract. Proliferation of epithelial cells was completely inhibited by dexamethasone as low as 30 nM. On the other hand, fibroblast-like stromal cells released by trypsin digestion required a plastic substratum coated with calf serum or fibronectin, and proliferated in Eagle's minimum essential medium supplemented with cholera toxin, bovine pituitary extract, dexamethasone, and bovine serum albumin. Epidermal growth factor and insulin had negligible effect on proliferation of stromal cells. Physiological concentrations of dihydrotestosterone and estradiol showed no effect on proliferation of both types of cells.
大鼠前列腺组织的不同酶消化程序以及独特的促有丝分裂因子组合,使得在不预先分离这两种细胞类型的情况下,实际上能够培养出纯净的上皮细胞和基质细胞群体。通过胰蛋白酶和胶原酶从成年大鼠腹侧前列腺解离的角蛋白阳性上皮细胞,在添加了表皮生长因子、胰岛素、霍乱毒素和牛垂体提取物的WAJC 404培养基中增殖。低至30 nM的地塞米松可完全抑制上皮细胞的增殖。另一方面,通过胰蛋白酶消化释放的成纤维细胞样基质细胞需要涂有小牛血清或纤连蛋白的塑料基质,并在添加了霍乱毒素、牛垂体提取物、地塞米松和牛血清白蛋白的伊格尔最低限度基本培养基中增殖。表皮生长因子和胰岛素对基质细胞的增殖影响可忽略不计。生理浓度的二氢睾酮和雌二醇对两种细胞的增殖均无影响。