诱导小鼠骨髓间充质干细胞向雄性生殖细胞的定向分化

目的探讨小鼠骨髓间充质干细胞是否能够在体外被诱导发生向雄性生殖细胞方向的分化。方法从雄性小鼠骨髓中分离能够长期贴壁生长的细胞,并鉴定其是否为间充质干细胞。对分离的细胞进行生殖细胞特异性报告基因标记(stra-8-GFP)。采用视黄酸诱导标记的细胞发生向生殖细胞方向的分化。通过观察报告基因表达和生殖细胞相关基因mRNA表达情况确定是否发生了分化。结果从小鼠骨髓中分离到的贴壁生长的细胞表达间充质干细胞的表面标志CD90、CD44、CD105和Sca-1;细胞在体外可以被诱导分化为成骨、成软骨及成脂肪细胞。报告基因标记的间充质干细胞在被视黄酸诱导2d后开始表达绿色荧光蛋白和生殖细胞相关基因Mvh、Fragilis和Stella的mRNA。未经视黄酸诱导的细胞不表达绿色荧光蛋白和生殖细胞相关基因。结论小鼠骨髓间充质干细胞在体外可以被视黄酸诱导发生向雄性生殖细胞方向的分化。

Study on differentiation of mesenchymal stem cells isolated from mouse bone marrow into male germ cells

Objective To investigate if mouse bone marrow derived-mesenchymal stem cell can be differentiated into male germ cell in vitro.Methods Mesenchymal stem cells were isolated from mouse bone marrow and were identified by surface markers and differentiation study in vitro.Germ cell specific reporter gene(stra-8-GFP) was transfected into the identified cells with Lipofectin 2000;Retinoic acid was used to induce the transfected cells to differentiate into germ cell.The germ cell differentiation was evaluated by expression of reporter gene and germ cell specific genes.Results The isolated cells had positive expression of CD90,CD44,CD105,Sca-1,similar with mesenchymal stem cells,and could be differentiated into osteocytes,chondrocytes and adipocytes in vitro.After being induced by retinoic acid for two days,the expression of GFP and germ cell specific gene mRNA were detected in the stra-8-GFP transfected cells.Conclusion Mouse bone marrow-derived mesenchymal stem cells could be differentiated into germ cells in vitro under induction of retinoic acid.