论文部分内容阅读
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。
Objective To investigate whether mouse bone marrow mesenchymal stem cells can be induced to differentiate into male germ cells in vitro. Methods The long-term adherent cells were isolated from the bone marrow of male mice and identified as mesenchymal stem cells. Isolated cells were germ-cell-specific reporter gene-tagged (stra-8-GFP). Retinoid-induced labeling of cells occurs in the direction of germ cell differentiation. Differentiation was confirmed by observation of reporter gene expression and germ cell-related gene mRNA expression. Results Adherent cells isolated from mouse bone marrow expressed mesenchymal stem cell surface markers CD90, CD44, CD105 and Sca-1; cells were differentiated into osteogenic, chondrogenic and adipocyte in vitro. Reporter-labeled mesenchymal stem cells began to express green fluorescent protein and germ cell-related genes Mvh, Fragilis and Stella mRNA 2d after induced by retinoic acid. Cells not induced by retinoic acid do not express green fluorescent protein and germ cell-related genes. Conclusion Mice bone marrow mesenchymal stem cells can be induced by retinoic acid to differentiate into male germ cells in vitro.