母体来源人胎盘间充质细胞的分离培养及其分化能力的研究

Isolation and Culture of Human Maternal Placenta Original Mesenchymal Stem Cells and Identification of its Differentiated Potential into Adipocytes and Osteoblasts

目的探讨胎盘母体来源间充质细胞的分离、培养方法及其向脂肪和成骨诱导分化潜能。方法采用酶消化获得母体来源胎盘间充质干细胞;体外扩增后,利用流式细胞仪检测其表面标志物;进行诱导分化后,采用油红O及茜素红染色鉴定其向脂肪和成骨诱导分化潜能。结果体外培养的母体来源胎盘间充质干细胞呈长梭形,细胞形态均一;细胞表面标志鉴定:CD73、CD90和CD105呈阳性表达,而CD14、CD34、CD45和HLA-DR呈阴性表达;细胞诱导分化后,经油红O、茜素红染色证实其可分化为脂肪细胞和成骨细胞。结论建立了母体来源胎盘间充质干细胞的分离培养方法;证实其具有成脂、成骨分化潜能,有望成为细胞治疗及组织工程更为理想的种子细胞。

Objeetive To search the suitable isolation and culture method of human maternal placenta original mesenchymal stem eells, and identify their differentiated potential into adipocytes and osteoblasts. Methods The mesenehymal stem cells were isolated from human maternal placenta by digested with collagenase; the expression of the specific cell surface marks was examined by flow cytometry; and the differentiated potential into adipocytes and osteblasts were identified by Oil Red O staining and Alizarlin Red staining. Results The human maternal placenta original mesenchymal stem cells exhibited homogeneous spindle shape; and they were positive expressed the eell surface marks of CDv3, CDg0 and CD105, but they negative expressed the cell surface marks of CD^4, CD34, CD45 and HLA-DR. After induced in differentiation, the cells were positive stained by Oil Red O staining and Alizarlin Red staining, which were the markers of adipocyte and osteoblast. Conclusion The isolation and culture methods of human matemal placenta original mesenehymal stem cells were established; the differentiated potential of human matemal placenta original mesenchymal stem cells into adipoeytes and osteoblasts were confirmed, which may provide an ideal alternative source for cell therapy and tissue engineering.