以EdU体外标记人脐带间充质干细胞:5,10μmol/L是其最适浓度  

EdU-labeled human umbilical cord mesenchymal stem cells in vitro:5 and 10μmol/L are the optimal concentrations of EdU

作　　者：王武[1] 张飞[1] 李贵才[1] 齐宝军[1] 武忠炎[1] 曹鹏[1] 

机构地区：[1]新疆医科大学第五附属医院骨科,新疆维吾尔自治区乌鲁木齐市830001

出　　处：《中国组织工程研究》2015年第32期5167-5171,共5页Chinese Journal of Tissue Engineering Research

基　　金：新疆医科大学科研创新基金项目(XYDCX2014167)~~

摘　　要：背景:Ed U为新一代细胞核标记物,目前对此标记的研究较少。目的:采取Ed U标记人脐带间充质干细胞,筛选出Ed U标记人脐带间充质干细胞的最适浓度。方法:采用组织块法分离、纯化及传代培养人脐带间充质干细胞,倒置显微镜观察其形态及生长情况,流式细胞仪鉴定细胞表面标志物,并进行成脂诱导分化能力鉴定。分别采用5,10,20,50,100μmol/L浓度的Ed U标记脐带间充质干细胞24 h,选择标记率最高的优化浓度,绘制细胞增殖曲线。结果与结论:倒置显微镜下观察细胞为贴壁生长,形态为长梭形,且EdU标记后的细胞形态与其一致,流式细胞仪检测CD44呈阳性表达,成脂诱导后具有向脂肪细胞分化的能力。5,10μmol/L Ed U的标记率最高,两种浓度标记的人脐带间充质干细胞生长曲线未见明显差异,故5,10μmol/L是体外标记人脐带间充质干细胞的最适浓度。BACKGROUND：EdU is a new nuclear marker, and currently, it is rarely reported. OBJECTIVE： To determine the optimal concentration of EdU to label human umbilical cord mesenchymal stem cels. METHODS： Human umbilical cord mesenchymal stem cels were isolated, purified and subcultured. Cel morphology and growth were observed under inverted microscope. Flow cytometry was used to identify cel surface markers, as wel as adipogenic identification. EdU at concentrations of 5, 10, 20, 50, 100 μmol/L was used to label human umbilical cord mesenchymal stem cels for 24 hours. The optimal concentration that resulted in the highest labeling efficiency was selected, and then cel proliferation curve was drawn. RESULTS AND CONCLUSION：Under the inverted microscope, cels grew adherently in a long spindle shape, and EdU-labeled cels had the same morphology. Flow cytometry showed that cels were positive for CD44, and had adipogenic differentiation ability. When the concentration of EdU was 5 and 10 μmol/L, the labeling efficiency was the highest, indicating that 5 and 10 μmol/L are the optimal concentrations of EdU to label human umbilical cord mesenchymal stem cels.

关 键 词：干细胞 脐带脐血干细胞 细胞示踪 脐带间充质干细胞 EDU 组织块法 

分 类 号：R394.2[医药卫生—医学遗传学]