体外诱导骨髓间充质干细胞定向分化为软骨细胞的实验研究  

In Vitro Induction of Directional Differentiation of Bone Marrow Mesenchymal Stem Cells towards Chondrocytes

作　　者：谭荣邦 史宏灿 

机构地区：[1]扬州大学临床医学院胸外科,江苏扬州225001

出　　处：《中国胸心血管外科临床杂志》2013年第3期325-328,共4页Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

基　　金：国家自然科学基金(30672080;30972968;81170014);江苏省"六大人才高峰"高层次人才基金(2009128);江苏省"青蓝工程"学术带头人基金(20081215)~~

摘　　要：目的体外分离培养、扩增骨髓间充质干细胞(mesenchymal stem cells,MSCs),诱导其定向分化为软骨细胞,为MSCs应用于临床、构建组织工程气管软骨提供实验依据。方法采用全骨髓培养+贴壁纯化法分离培养MSCs,流式细胞仪鉴定MSCs,以转化生长因子β1(transforming growth factorβ1,TGF-β1)、胰岛素样生长因子1(insulin-like growth factor 1,IGF-1)作为主要的诱导因子诱导MSCs定向分化为软骨细胞。21 d后用Ⅱ型胶原免疫细胞化学染色法鉴定诱导出的细胞,在光学显微镜和电子显微镜下观察MSCs诱导前、后的细微和超微结构改变。结果诱导21 d后Ⅱ型胶原免疫细胞化学染色呈阳性。光学显微镜和电子显微镜检查显示:MSCs呈梭形,诱导后细胞增大,呈多边形、星形、三角形。透射电子显微镜显示:诱导后细胞器丰富,胞核增大,核仁增多。结论 MSCs诱导分化为软骨细胞的超微结构表现为细胞器丰富、胞核增大、核仁增多,提示细胞分化、代谢功能旺盛。Objective To isolate, culture and expand bone marrow mesenchymal stem cells （MSCs） in vitro, induce MSCs to differentiate directionally towards chondrocytes, and provide experimental basis for clinical application of MSCs and construction of tissue engineering tracheal cartilage. Methods Cultured MSCs were isolated from bone marrow of Sprague-Dawley rats, purified using adherence separation, and identified by flow cytometry analysis. Transforming growth factor β1 （ TGF-β 1 ） and insulin-like growth factor 1 （ IGF- 1 ） were used as main induction factors to induce MSCs to differentiate directionally towards chondrocytes. The expression of collagen typeⅡ was evaluated by immunocytochemical staining 21 days after induction. Light microscope and electron microscope were used to observe tiny and ultrastructural changes of the cells before and after induction. Results The expression of collagen type Ⅱ was positive by immunocy- tochemical staining 21 days after induction. MSCs were fusiform before induction under light microscope and electron microscope. After induction, the cells became larger, polygon, star-shaped or triangular. Transmission electron microscope showed that the cells had abundant organelles, larger nuclei and more nucleoli after induction. Conclusion Abundant organelles, larger nuclei and more nucleoli are the ultrastructure changes of chondrocytes differentiated from MSCs, indicating that the cells are active in differentiation and metabolism.

关 键 词：间充质干细胞 软骨分化 超微结构 Ⅱ型胶原 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]