人胎盘绒毛膜间充质干细胞分离培养的新方法

摘要

To establish an effective and easy method of isolating and cultivating human placental chorionic-derived mesenchymal stem cells (hpcMSCs).Human placental chorion were isolated by hand held electric homogenizer and enzyme digestion.The morphol-ogy of hpcMSCs was observed under the inverted microscope.The cell growth curve was measured by CCK-8 method.The third gen-eration of hpcMSCs were detected with flow cytometer,and incubated in osteogenesis and adipogenesis.Osteogenous potential was as-sessed by alizarin red staining.Adipogenic potential was evaluated by Oil red O staining.The adherent cells showed fibroblast-cell-like shape.CD73, CD105 and CD90 membrane antigens could be detected in the third generation of hpcMSCs,but CD34, CD45, CD14, CD19 and HLA-DR membrane antigens were not detected.Following induction, alizarin red staining and Oil red O staining produced a strong reaction in cells.The whole isolate and cultivation is an effective and easy method to obtain a largely amount of prima-ry hpcMSCs.It may represent a plentify stem cell source for the regenerative medicine.%建立一种简便、快捷的人胎盘绒毛膜间充质干细胞(human placental chorionic -derived mesenchy-mal stem cells, hpcMSCs)分离培养方法.用手持电动匀浆器处理人绒毛膜,经酶消化后接种培养,用倒置显微镜观察细胞形态,CCK-8测定细胞生长曲线,流式细胞术检测细胞表面标记,成脂、成骨诱导分化试剂盒鉴定细胞的分化潜能.获得的人胎盘绒毛膜间充质干细胞为成纤维细胞样细胞,呈平行排列或漩涡状生长,高表达CD73、CD105、CD90,而CD34、CD45、CD14、CD19、HLA-DR呈阴性,生长曲线为" S"型,经成骨或成脂诱导后,茜素红染色或油红O染色呈阳性.符合国际细胞治疗学会认可的间充质干细胞标准.建立了人胎盘绒毛膜间充质干细胞的分离培养新方法,结合了组织块法和酶消化法的优点,为绒毛膜间充质干细胞的应用提供基础.