人骨髓间充质干细胞体外向软骨细胞分化的实验研究  

Research on differentiation of human bone marrow-derived mesenchymal stem cells into chondrocytes in vitro

作　　者：吴家昌[1] 杨彤涛[1] 许啸[1] 文艳华[1] 韩建伟[1] 范清宇[1] 马保安[1] 

机构地区：[1]第四军医大学唐都医院骨肿瘤研究所,西安医学硕士研究生710038

出　　处：《医学研究生学报》2010年第2期128-132,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金(30672143)

摘　　要：目的创伤等因素造成的关节软骨的损伤和退变是骨科常见和难以解决的问题之一。干细胞尤其是骨髓来源干细胞的研究促进了组织工程在骨关节领域的发展。文中研究体外单层培养条件下,成人骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSC)经诱导定向分化软骨细胞可行性。方法用全血贴壁法分离培养BMSC,并传代扩增后,用单层培养法在软骨诱导剂下进行诱导培养21 d,以常规培养液培养的BMSC作为阴性对照。光镜下观察诱导细胞形态学的改变,苏木素-伊红(HE)染色、甲苯胺蓝染色、Ⅱ型胶原蛋白免疫组化等方法检测诱导细胞的分泌功能。结果BMSC经21 d的单层诱导培养后,细胞形态由长梭形向多角形类圆形转变,细胞爬片甲苯胺蓝染色呈明显异染,Ⅱ型胶原蛋白免疫细胞化学检测阳性,对照组阴性。结论成人BMSC在单层培养条件下可成功诱导分化为软骨细胞。Objective Lesions and degradations of articular cartilage that caused by many factors such as trauma are commonly encountered knotty medical problem. The research of stem cells （i. e. , bone marrow-derived steml cells） have given rise to the new field of tissue engineering. This article is to investigate the feasibility of inducing human bone marrow mesenchymal stem cells （hBMSC） into chondrocytes in monolayer culture in vitro. Methods The bone marrow mesenchymal stem cell （BMSC） obtained in bone marrow aspirates from adult were isolated by the whole marrow-adherence way from the mononuclear cells, and then were induced into chondrocyte in monolayer by a chemically defined medium including TGF-β1, insulin , dexamethasone and so on. BMSC were cultured in monolayer in the conventional culture medium as negative control. After 21-day＇s chondrogenic inducement, the cells were analysed by histological observation, staining with haematoxylin-eosin and toluidine blue, immunohistochemistry evaluation with the monoclonal antibody specific to type Ⅱ collagen. Results BMSC were induced in monolayer condition for 21 days by defined medium were metachromasia by toluidine blue and could be identified by positive staining for type Ⅱ collagen. The control groupe was not. Conclusion The hBMSCs can be induced into chondrocytes in monolayer by the chemically defined medium.

关 键 词：人骨髓间充质干细胞 体外诱导 分化 软骨细胞 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]