大鼠骨髓间充质干细胞与睾丸支持细胞共培养的生物学特征

Biological characteristics of co-culture of mesenchymal stem cells and Sertoli cells in mice

目的研究骨髓间充质干细胞（mesenchymal stem cells，MSCs）在体外与睾丸Sertoli细胞共培养，模拟体内生精微环境条件下的生物学特征。方法分离、纯化、培养大鼠骨髓MSCs、睾丸Sertoli细胞（SCs）、精原干细胞（SSCs），设SCs组、SCs饲养层＋SSCs组、MSCs组、SCs饲养层＋MSCs组，均用条件培养基培养，观察各组细胞形态、存活及生长情况，检测各组碱性磷酸酶（AKP）表达情况。结果与单独培养的MSCs和SCs相比较，共培养之MSCs和sCs存活时间延长，MSCs源性细胞收缩变圆形，折光性增强，可见由细胞间桥连接的圆形双细胞或四细胞，与SSCs一样呈碱性磷酸酶阳性表现。结论在体外与睾丸Sertoli细胞共培养，模拟体内生精微环境的条件下，MSCs能长期存活，并获得SSCs的部分生物学特征，有可能转分化为生精细胞。

Objective To study the biological characteristics of co-culture of bone marrow mesenchyrnal stem cells （MSCs） and Sertoli cell layer in vitro as a mimicked microenvironment of spermatogenesis in mice. Methods The MSCs,Sertoli cells （SCs） and spermatogonia stem cells （SSCs） from Wistar mice aging 1d,3d and 311-d after birth were isolated, purified and divided into 4 groups as follows： SCs,SCs＋ SSCs,MSCs and SCs＋ MSCs. All of them were cultured in the conditioned medium. The morphology, survival and growth status were observed, and the expression of alkaline phosphatase was detected simuhaneously. Results Compared isolated MSCs with SCs culture group, the survival time of co culture of MSCs with SCs was prolonged part of the cells of MSCs shrank to round shape with a high refraction,The paired round cells or quadrifid round-cells were observed with alkaline phosphatase positive reaction. Conclusions During the bone marrow mesenchyrnal stem cells co- cultured in vitro with Sertoli cells to mimic microenvironment of spermatogenesis, the MSCs can be survived for a long time and obtained some characteristics of S,SCs,which can probably be transformed in to spermatogenic cells.