小鼠脐带间充质干细胞的体外诱导分化

Isolation and differentiation in vitro of mouse umbilical cord mesenchymal stem cells

目的分离扩增小鼠脐带间充质干细胞(mouse umbilical cord mesenchymal stem cells,m UCMSCs)探讨其是否可诱导成软骨、脂肪和成骨细胞。方法通过贴壁培养法将m UCMSCs体外分离、扩增、纯化,倒置显微镜下观察细胞的形态特征,运用流式细胞仪检测分析细胞的抗原标志表达进行鉴定。运用诱导培养液对分离的m UCMSCs分别定向诱导培养为软骨、脂肪和成骨细胞。结果运用组织贴块培养法可从新鲜脐带中分离到贴壁生长的成纤维样细胞,这些细胞高表达CD29、CD90和CD105,低表达CD34。成软骨诱导后阿新兰染色呈蓝色;成脂诱导后油红O染色,出现红色脂滴;茜红素染色成骨诱导的m UCMSC,可见红色结节。结论贴壁培养法分离培养所获得的m UCMSCs在体外可诱导分化为软骨、脂肪和成骨细胞。

Objective To isolate,culture and identify mouse umbilical cord mesenchymal stem cells（ m UCMSCs）and to study whether they can be induced to differentiate into adipocytes,chondrocytes and osteoblasts. Methods The m UCMSCs were isolated and expanded by adherent culture from fresh mouse umbilical cord. The morphological characteristics of the resulting cells were observed under inverted phase contrast microscope,and their expression of mesenchymal surface markers was identified and analyzed by flow cytometry. Then m UCMSCs were induced to differentiate into chondrocytes,adipocytes and osteoblasts in vitro. Results Fibroblast-like cells could be isolated from the fresh umbilical cord by adherent culture. These adherent cells highly expressed mesenchymal markers including CD29,CD90 and CD105 while low expression of CD34. The cells were successfully induced to differentiate into chondrocytes,adipocytes and osteoblasts.Conclusions The m UCMSCs isolated from fresh mouse umbilical cord by adherent culture have potential of differentiation into chondrocytes,adipocytes and osteoblasts.