骨髓间充质干细胞定向诱导成类软骨细胞的实验研究

目的　体外诱导骨髓间充质干细胞 (MSCs)向软骨细胞表型分化 ,探讨其分化的条件 ,为其作为软骨组织工程种子细胞提供实验基础。方法　取第二代成年大鼠骨髓间充质干细胞 ,实验组用无血清培养液诱导 ;对照组用含 10 %胎牛血清的培养液自然分化。Ⅱ型胶原免疫组化、甲苯胺蓝染色、透射电镜检测其分化情况。统计学分析不同时间、不同接种密度的软骨分化率。结果　诱导后的MSCs具有软骨细胞特点 ,不同诱导时间、不同接种密度的Ⅱ型胶原免疫组化阳性率有显著差异。诱导 14d为最佳 ,诱导率与细胞接种密度成正相关。结论　成年大鼠骨髓间充质干细胞在无血清培养基能向软骨细胞方向转化。

Experimental Study of Differentiation of Marrow Mesenchymal Stem Cell into Chondrocyte-like Cells

Objective Induction of bone marrow mesenchymal stem ce ll s (MSCs) of adult rat in vitro was performed to explore the possibility of MSCs being cartilage seed cell in tissue engineering. Methods MSCs of the second passage of adult rat were divided into test group induced by free -serum HG-DMEM and control group induced by HG-DMEM with 10% fetal bovine se rum respectively. Collagen type Ⅱ immunohistochemical staining, toluidine blue staining, transmisson electron microscope were performed to detect differentiati on. Statistical analysis of the MSCs differentiation was deployed under various induction time and various plating density. Results Induced M SCs have characteristics of the chondrocyte, the positive percentage of collagen type Ⅱ immunohistochemical staining was different statistically between variou s induction time and various plating density. 14 days induction in culture is th e best time, induction rate is positively related to plating density. C onclusion Free-serum HG-DMEM culture can induce MSCs differentiating into chondrocyte-like cells.