大鼠脂肪组织源间充质干细胞的培养及诱导分化  

Culture and Inducing Differentiation of Mesenchymal Stem Cells Derived from Adipose Tissue of Rat

作　　者：郭勇[1] 张西正[1] 魏严[1] 郭春[1] 李瑞欣[1] 徐晓莹[1] 张永红[1] 

机构地区：[1]军事医学科学院卫生装备研究所,天津300161

出　　处：《航天医学与医学工程》2008年第6期477-481,共5页Space Medicine & Medical Engineering

基　　金：天津市自然科学基金重点项目(06YFJZJC02000);全军医药卫生科研基金面上项目(06MA344);国家自然科学基金面上项目(30700156)

摘　　要：目的体外培养脂肪组织源间充质干细胞,诱导该细胞向成骨细胞和心肌细胞分化,为骨和心肌组织工程提供种子细胞。方法取Wistar大鼠脂肪组织,以胰蛋白酶消化分离出脂肪间充质干细胞。第三代细胞用于鉴定细胞表面抗原,同时以成骨诱导培养液诱导细胞,并以10μmol/L5-氮胞苷处理细胞24h,然后培养,用免疫组化法和组织化学法检测这些细胞。结果所分离细胞90%以上表达CD44、CD13、CD29抗原,有些细胞表达肌节型肌动蛋白和结蛋白;经成骨诱导液诱导14d的细胞表达碱磷酸酶,21d后表达骨钙蛋白并有少量胞外钙沉积;经10μmol/L5-氮胞苷处理的细胞均表达NKx2.5、肌节型肌动蛋白和连接蛋白Cx43。结论本实验所培养的细胞为脂肪组织源性间充质干细胞,能向成骨细胞和心肌样细胞分化,可为骨和心肌组织工程提供种子细胞。Objective To provide seed cells for bone and cardiac tissue engineering, adipose tissue-derived mesenchymal stem cells cultured in vitro and induced differentiation into osteoblast and cardiomyocyte-like cells. Methods The mesenchymal stem cells were separated and cultured from the adipose tissue of Wistar rats with typsin digestion. The third passage cells were used to be induced differentiation and detected the surface antigens. Some cells were cultured with osteoinductive medium,the other cells were cultured with 10 μmol/L 5-Azacytidine for 24 h,then the cells were detected by immunohistochemical and histochemical method. Results More than 90% separated cells expressed CD44, CD13 and CD29 antigens. Some cells expressed α-sacromeric actin and desmin. Induced with osteoinductive medium for 14 d, cells expressed alkaline phosphatase. Induced for 21 d, cells expressed osteocalcin and a little extracelluiar calcium sediment appeared. After 14 d cultured with 10 μmol/L 5-Azacytidine,the cells were found to express NK×2.5 α-sacromeric actin and C×43. Conclusion Mesenchymal stem cells can be obtained from adipose tissue by digestion method. These cells can be differentiated into osteoblast and cardiomyocyte-like cells to provide seed cells for bone and cardiac tissue engineering.

关 键 词：脂肪组织 间充质干细胞 分化 成骨细胞 心肌细胞 组织工程 

分 类 号：Q813.1[生物学—生物工程]