Iguchi T, Uchima F D, Bern H A
In Vitro Cell Dev Biol. 1987 Aug;23(8):535-40. doi: 10.1007/BF02620970.
Normal mouse vaginal epithelial cells isolated from ovariectomized ca. 35-d-old BALB/cCrgl mice were grown in primary culture using collagen gel matrix and a serum-free medium composed of a 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 (D:H) medium supplemented with insulin (IN), epidermal growth factor (EGF), cholera toxin, transferrin, and bovine serum albumin V (BSA). Three-dimensional cellular outgrowths occurred inside the collagen gel matrix. The contribution of each factor to cell growth was examined by individual addition to the basic D:H medium and by individual deletion from the complete serum-free medium. When added individually, only IN promoted growth. Deletion of IN from the complete serum-free medium markedly diminished growth; deletion of EGF or BSA slightly diminished growth. When horse, fetal bovine, or chicken serum was added to the basal D:H medium, only with increasing doses of horse serum was there enhanced cell growth. The effect of 17 beta-estradiol and diethylstilbestrol on the growth of cells was also tested, using a suboptimal medium of D:H supplemented with BSA and IN, or a minimal medium supplemented with IN alone. During the 8-d time period, addition of estrogen did not enhance cell growth in either medium. To date, we have been unable to demonstrate a mitogenic effect of estrogen; rather a dose-dependent inhibition of proliferation is seen.
从约35日龄卵巢切除的BALB/cCrgl小鼠分离出的正常小鼠阴道上皮细胞，使用胶原凝胶基质和由杜氏改良 Eagle 培养基与哈姆氏F12（D:H）培养基按1:1比例混合并添加胰岛素（IN）、表皮生长因子（EGF）、霍乱毒素、转铁蛋白和牛血清白蛋白V（BSA）组成的无血清培养基进行原代培养。胶原凝胶基质内出现了三维细胞生长。通过单独添加到基础D:H培养基以及从完全无血清培养基中单独去除每种因子，来检测各因子对细胞生长的作用。单独添加时，只有IN能促进生长。从完全无血清培养基中去除IN会显著降低生长；去除EGF或BSA会使生长略有降低。当向基础D:H培养基中添加马血清、胎牛血清或鸡血清时，只有随着马血清剂量增加细胞生长才会增强。还使用添加了BSA和IN的次优D:H培养基或仅添加IN的基本培养基，测试了17β-雌二醇和己烯雌酚对细胞生长的影响。在8天时间段内，在两种培养基中添加雌激素均未增强细胞生长。迄今为止，我们未能证明雌激素有促有丝分裂作用；相反，观察到的是剂量依赖性的增殖抑制。
