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小鼠脐带间充质干细胞的体外诱导分化
引用本文:朱慧,叶莉,何洁,余璞,王金祥,王强,赵晶,庞荣清,白杰英. 小鼠脐带间充质干细胞的体外诱导分化[J]. 实验动物与比较医学, 2015, 23(6): 622-627
作者姓名:朱慧  叶莉  何洁  余璞  王金祥  王强  赵晶  庞荣清  白杰英
作者单位:成都军区昆明总医院云南省干细胞工程实验室,军事医学科学院实验动物中心,成都军区昆明总医院云南省干细胞工程实验室,成都军区昆明总医院云南省干细胞工程实验室,成都军区昆明总医院云南省干细胞工程实验室,成都军区昆明总医院云南省干细胞工程实验室,成都军区昆明总医院云南省干细胞工程实验室,军事医学科学院实验动物中心,成都军区昆明总医院云南省干细胞工程实验室
基金项目:云南省基金项目(2011HB050, 2013DA004),十二五重大项目(2011ZXJ09201-031),国家自然科学基金(31272385),北京市自然科学基金(5152023)
摘    要:目的分离扩增小鼠脐带间充质干细胞(mouse umbilical cord mesenchymal stem cells,mUCMSCs)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mUCMSCs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mUCMSCs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mUCMSC,可见红色结节。结论 贴壁培养法分离培养所获得的mUCMSCs在体外可诱导分化为软骨、脂肪和成骨细胞。

关 键 词:小鼠;脐带间充质干细胞;细胞培养;成软骨诱导;成脂诱导;成骨诱导
收稿时间:2015-06-01
修稿时间:2015-06-25

Isolation and differentiation in vitro of mouse umbilical cord mesenchymal stem cells
ZHU Hui,YE Li,HE Jie,YU Pu,WANG Jin-xiang,WANG Qiang,ZHAO Jing,PANG Rong-qing and BAI Jie-ying. Isolation and differentiation in vitro of mouse umbilical cord mesenchymal stem cells[J]. Laboratory Animal and Comparative Medicine, 2015, 23(6): 622-627
Authors:ZHU Hui  YE Li  HE Jie  YU Pu  WANG Jin-xiang  WANG Qiang  ZHAO Jing  PANG Rong-qing  BAI Jie-ying
Affiliation:Key Laboratory of Stem Cells and Regenerative Medicine, PLA Kunming General Hospital of Chengdu Military Region, Kunming 650032, China;Kunming Medical University Clinical College in Kunming General Hospital Kunming 650032;Laboratory Animal Center of the Academy of Military Medical Sciences, Beijing 100071;Key Laboratory of Stem Cells and Regenerative Medicine, PLA Kunming General Hospital of Chengdu Military Region, Kunming 650032, China;Key Laboratory of Stem Cells and Regenerative Medicine, PLA Kunming General Hospital of Chengdu Military Region, Kunming 650032, China;Kunming Medical University Clinical College in Kunming General Hospital Kunming 650032;Key Laboratory of Stem Cells and Regenerative Medicine, PLA Kunming General Hospital of Chengdu Military Region, Kunming 650032, China;Key Laboratory of Stem Cells and Regenerative Medicine, PLA Kunming General Hospital of Chengdu Military Region, Kunming 650032, China;Key Laboratory of Stem Cells and Regenerative Medicine, PLA Kunming General Hospital of Chengdu Military Region, Kunming 650032, China;Key Laboratory of Stem Cells and Regenerative Medicine, PLA Kunming General Hospital of Chengdu Military Region, Kunming 650032, China;Laboratory Animal Center of the Academy of Military Medical Sciences, Beijing 100071
Abstract:Objective To isolate, culture and identify mouse umbilical cord mesenchymal stem cells(mUCMSCs)and to study whether they can be induced to differentiate into adipocytes, chondrocytes and osteoblasts.Methods The mUCMSCs 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 mUCMSCs 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 mUCMSCs isolated from fresh mouse umbilical cord by adherent culture have potential of differentiation into chondrocytes, adipocytes and osteoblasts.
Keywords:Mouse  Umbilical cord mesenchymal stem cells  Cell culture  Chondrocytes induction  Adipocytes induction  Osteoblast induction
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