卵泡刺激素和表皮生长因子对小鼠精原细胞增殖的影响

利用生殖细胞-体细胞体外无血清共培养模型研究了卵泡刺激素(FSH)和表皮生长因子(EGF)对小鼠A型精原细胞增殖的影响。精原细胞在ITS培养液(添加胰岛素、转铁蛋白和亚硒酸钠的DMEM)中培养24h后进行c-kit免疫细胞化学鉴定和EGF及其受体(EGFR)免疫细胞化学检测,72h后测定其形成集落数的情况。结果表明:ITS培养液能维持生殖细胞的活性,增殖细胞核抗原(PCNA)的表达增高。A型精原细胞呈c-kit阳性,EGF和EGFR主要表达于精原细胞。单独的FSH(1～100ng/ml)或EGF(1～10ng/ml)显著促进精原细胞集落数的增加。此外,EGF(0.1ng/ml)联合FSH(10ng/ml)具有加性效应,但更高剂量的EGF(1～10ng/ml)则降低了FSH的刺激作用。结果说明FSH可联合适量的EGF促进精原细胞的增殖。

Effects of Follicle-stimulating Hormone and Epidermal Growth Factor on Proliferation of Mouse Spermatogonial Cells

A serum-free germ-somatic cell coculture model was established to evaluate the effects of follicle-stimulating hormone (FSH) and epidermal growth factor (EGF) on proliferation of mouse type A sper-matogonial cells. Immunocytochemical staining for c-kit,EGF and EGF receptor (EGFR) were performed after 24 h culture and cell proliferation was assessed after 72 h culture. Results showed that ITS medium supplemented with insulin,transferring and selenite could maintained spermatogonial cell survival,with increased expression of prolif-erating cell nuclear antigen. Type A spermatogonial cells manifested positive c-kit staining,and both EGF and EGFR were mainly expressed in spermatogonial cells. FSH (1-100 ng/ml) or EGF (1-10 ng/ml) treatment significantly increased spermatogonial colony number. Furthermore,EGF (0.1 ng/ml) could synergize with 10 ng/ml FSH to increase the colony number,but inhibited FSH-stimulated effect at 1-10 ng/ml. The above results indicated that EGF could synergize with FSH to promote proliferation of spermatogonial cells.