【摘要】 为了探究表皮生长因子(EGF)对体外培养的精原干细胞增殖的调控作用及其作用机制.应用不连续Percoll梯度液和选择性贴壁法分离纯化精原干细胞,c-kit细胞免疫组化鉴定细胞,MTT法研究EGF对精原干细胞增殖的效应,再加入JAK-STAT信号通路特异性抑制剂AG490,探究EGF对精原干细胞增殖作用的可能机制.c-kit细胞免疫组化结果显示分离得到细胞为精原干细胞;MTT结果显示各实验组比对照组细胞数量均有显著增多(P<0.01);与对照组相比,加入AG490组的活细胞数有显著下降(P<0.01).实验结果表明EGF能够促进精原干细胞的增殖,并且可以通过JAK-STAT信号通路起作用.更多还原

【Abstract】 To study the stimulative effects of epidermal growth factor (EGF) on the proliferation of the spermatogonial stem cells and its mechanisms.Spermatogonial stem cells were isolated by discontinuous Percoll gradient centrifugation and selective adherence.The Anti c-kit antibody was applied to identify spermatogonial stem cells.The spermatogonial stem cells were cultured with EGF at different concentrations in vitro and the quantity of live cells was detected by MTT assay.EGF and AG490 (janus kinase inhibitor,JAK-STAT) were added respectively to the medium and the percentage of the live cells was determined by MTT assay after incubation.The cells which we harvested were spermatogonial stem cells.EGF significantly enhanced the number and percentage of cell survival in a dose-dependent manner (P < 0.01).Compared with EGF group,the percentage of cell survival in AG490 group reduced markedly (P < 0.01).The result of experiment indicates that EGF could promote spermatogonial stem cells proliferation obviously.The effect of EGF on the spermatogonial stem cells can be mediated by the activations of intracellular JAK-STAT system.更多还原