人脐带华通氏胶间充质干细胞的生物学特性及其向肌源性细胞分化的能力

Biological characteristics of human umbilical cord Wharton′s Jelly mesenchymal stem cells and their ability in differentiation to myogenic cells

目的探讨人脐带华通氏胶间充质干细胞(human umbilical cord Wharton′s Jelly mesenchymal stem cells,hUCMSCs)的生物学特性及其向肌源性细胞分化的能力。方法收集健康足月产胎儿脐带组织,采用酶消化法从华通氏胶中分离hUCMSCs,选取第3代对数生长期细胞,显微镜观察细胞形态,流式细胞术检测细胞免疫表型分子的表达,免疫荧光染色检测干细胞标记物及其体外向肌源性细胞的分化,荧光显微镜观察及流式细胞术检测携带绿色荧光蛋白(green fluorescent protein,GFP)的慢病毒转染hUCMSCs后GFP的表达。结果采用酶消化法获得可贴壁生长的大量hUCMSCs,其能在体外成功进行扩增培养。hUCMSCs可高表达CD29、CD44、CD73、CD90和CD105,而极低表达CD14、CD34、CD45和HLA-Ⅱ;能表达干细胞标记物Oct4和Sox2,且在体外特殊培养条件下表达骨骼肌干细胞标记物Pax7和Myo D;转染的hUCMSCs可稳定表达GFP,转染率高达50%~70%。结论人脐带华通氏胶组织存在间充质干细胞,且其具有向肌源性细胞分化的潜能。

Objective To investigate the biological characteristics of human umbilical cord Whalton′ s jelly mesenchymal stem cells（hUCMSCs）as well as their ability in differentiation to myogenic cells. Methods The umbilical cord tissue of healthy full-term fetuses,from which hUCMSCs were isolated by digestion with collagenase Ⅱ. The hUCMSCs of passage 3 in logarithmic growth period were observed for morphology by microscopy, analyzed for expression of cellular immunophenotypic molecules by flow cytometry,and for stem cell markers and the differentiation of the cells to myogenic cells in vitro by IFA. The expression of green fluorescent protein（GFP） in hUCMSCs transfected with lentivirus carrying GFP gene was observed by IFA and flow cytometry. Results A large quantity of adherent hUCMSCs were obtained,which could be cultured and proliferated in vitro. CD29,CD44,CD73,CD90 and CD105 were highly expressed,while CD14, CD34, CD45 and HLA-Ⅱ were lowly expressed, in hUCMSCs. Stem cell markers Oct4 and Sox2 were expressed,moreover,skeletal muscular stem cell markers Pax7 and MyoD were expressed under special culture condition in vitro,in hUCMSCs. GFP was stably expressed in hUCMSCs transfected with lentivirus,while the transfection rate was50%. Conclusion Mesenchymal stem cells existed in human umbilical cord Wharton′s Jelly, which showed the capacity of differentiation to myogenic cells.