摘要
建立一种简单、有效体外分离和培养精原干细胞(Spermatogonia stem cell,SSCs)的方法。采用酶消化6～8d新生小鼠睾丸制备睾丸细胞悬液后进行培养,碱性磷酸酶(AKP)染色和间接免疫荧光对培养的细胞进行鉴定;结果显示:SSCs培养3～5天可观察到细胞克隆的产生,AKP染色强阳性;间接免疫荧光显示GCNF阴性,oct-4、c-kit均呈阳性反应。由此可知以DMEM+15%胎牛血清+白血病抑制因子(1eukemia inhibitory factor,LIF)+L-谷氨酰胺等为培养基,成功建立了精原干细胞的体外培养体系。
To develop a simple and effective method by which spermatogonial stem cell （SSCs） can be isolated and cultured from mice testis. Enzymatic digestion was used to prepare suspension of SSCs from newborn mice testis , and then culture in vitro. Alkaline phosphorase （AKP） histochemical staining and indirect immunofluorescence staining were used to identify cultured cells. Cell clones could be observed after 3-4d of primary cell culture. The cultured cells possessed the biological behaviors of spermatogonial stem cell. AKP histochemical staining shows strong positive. Indirect immunofluorescence show that both Oct-4 and c-kit had positive staining, but GCNF was negative. A successful primary cell culture system for mouse spermatogonial stem cell was establisted, in which the cells cultured on DMEM medium supplemented with 15% FBS, LIF, L-glutamine .
出处
《石河子大学学报（自然科学版）》
CAS
2009年第4期470-473,共4页
Journal of Shihezi University(Natural Science)
关键词
精原干细胞
小鼠
原代培养
体外研究
Spermatogonial stem cell
Mice
Primary cell culture
In vitro
