睾丸提取液、EGF、雌激素对小鼠精原干细胞体外存活和增殖的效果

为探讨成年小鼠睾丸提取液（testicular abstract, TA）、表皮生长因子（epidermal growth factor, EGF）、雌二醇-17β(estradiol-17β, E2)对小鼠精原干细胞（spermatogonial stem cells, SSCs）体外存活及增殖的作用。用胶原酶-胰蛋白酶消化、差速贴壁法分离8 d小鼠的SSCs，培养于STO细胞饲养层上，培养液中分别添加不同浓度的TA、EGF和E2，观察SSCs的存活时间和增殖能力。结果显示，添加3种因子后，SSCs的平均存活时间与对照组相比虽无显著差异。但在20~40 ng/ml EGF和10~100 pg/ml E2组中SSCs的平均存活时间均明显延长，且在较长时间段内观察到了处于明显增殖期的细胞。提示20~40 ng/ml EGF和10~100 pg/ml E2对小鼠SSCs的体外存活和增殖具有一定良好作用。

Effects of Testicular tract, Epidermal Growth Factor and Estradiol-17B on the Maintenance of Mouse Spermatogonial Stem Cells in Vitro

To investigate the effects of testicular abstract (TA) from adult mice testes, epidermal growth factor (EGF) and Estradiol-17β (E2) on the survival and proliferation of mouse spermatogonial stem cells (SSCs) in vitro. The seminiferous epithelium of 8-day-old mouse was dissociated with collagenase-trypsin digestion. SSCs were enriched by differential plating and seeded on STO feeder layers in a compound media containing varied concentrations of TA, EGF and E2. The survival and proliferation of SSCs were investigated. The results showed that no significant difference was observed between the average survival times of SSCs in experiment groups and that in control groups after adding with the three factors. However, the survival times of SSCs in groups containing 20~40 ng/ml EGF and 10~100 pg/ml E2 were obviously extended, and distinct proliferation activities of SSCs were observed during a relatively long period. The results indicate that EGF of 20~40 ng/ml and E2 of 10~100 pg/ml were beneficial to the maintenance of mouse SSCs.