骨髓间充质干细胞定向诱导成类软骨细胞的实验研究(英文)

Experimental study on the chondroyte like differentiation of marrow mesenchym al stem cell

背景:用组织工程的方法修复软骨缺损克服了传统方法的诸多不足。目的:体外诱导骨髓间充质干细胞向软骨细胞表型分化,探讨其作为软骨组织工程种子细胞的可行性。设计:完全随机设计,对照实验研究。单位:哈尔滨医科大学组胚教研室及神经生物教研室。材料:Wistar大鼠清洁级动物6只,雌雄不限,来源于哈尔滨医科大学()附属第二医院实验动物中心,实验动物生产许可证号:SCXK(黑)20020002。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、甲苯胺蓝染色检测其分化情况。主要观察指标:软骨细胞的鉴定,不同诱导时间Ⅱ型胶原免疫组化阳性率的比较。结果:诱导后的MSCs具有软骨细胞的特点。结论:成年大鼠骨髓间充质干细胞在特定的培养基能向软骨细胞方向转化,作为软骨组织工程种子细胞具有可行性。

BACKGROUND:Tissue engineering method has been employed to recover cartilage de fect and overcome many traditional shortages. OBJECTIVE:In vitro marrow mesenchymal stem cells(MSCs) of adult rat was induce d to differentiate into chondrocyte phenotype so as to probe into the feasibilit y of MSCs to be cartilage seed cell in tissue engineering. DESIGN:Completely randomized design and controlled experimental study. SETTING:Department of Histology and Embryology,and Department of Neurobiology, Harbin Medical University MATERIALS:Six Wistar rats of either gender,cleanness grade,were provided by Ex perimental Animal Center,Affiliated Second Hospital,Harbin Medical University.Pe rmission number of experimental animal production was SCXK(black) 20020002. METHODS:The MSCs of the second generation adult rat was taken and divided into test group and control group.The test group was induced with serum free and con trol group was induced with 10%fetal calf serum.Collagen type Ⅱimmunohistochem ical staining, toluidine blue staining was performed to detect differentiation. MAIN OUTCOME MEASURES:Identification of chondrocyte,comparison of the positive rate of Collagen type Ⅱimmunohistochemical staining at different induction tim e point. RESULTS:Induced MSCs had identical characteristic to the chondrocyte. CONCLUSION:In vitro culture can induce MSCs to differentiate directly to chond rocyte like cell.The results suggest that it is feasible to use MSCs as seed ce lls in the cartilage tissue engineering.