体外微团培养兔骨髓间充质干细胞形成软骨细胞  

Inducing bone mesenchymal stem cells of rabbits into chondrocytes using the technology of micromass culture in vitro

作　　者：李斌[1] 张伟[1] 王健[1] 孙水[1] 吴帅[1] 

机构地区：[1]山东大学附属省立医院关节外科,济南250021

出　　处：《中国矫形外科杂志》2009年第23期1819-1822,共4页Orthopedic Journal of China

基　　金：山东省医药卫生科研计划项目(2005-97)

摘　　要：[目的]探讨兔骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)体外诱导形成软骨细胞的方法,以及转化生长因子-β1(transforming growth factorβ1,TGF-β1)、胰岛素样生长因子-Ⅰ(insulin-like growth factor-Ⅰ,IGF-Ⅰ)和碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)之间的相互作用。[方法]对兔骨髓间充质干细胞体外进行原代和传代培养,按加入诱导条件不同分为四组:A组:TGF-β1+bFGF;B组:TGF-β1+IGF-Ⅰ;C组:TGF-β1;D组:空白对照组。3周后分别做四唑盐(methyl thiazolyl tetrazolium,MTT)比色试验、糖胺聚糖(glycosaminoglycan,GAG)检测和免疫组织化学染色。[结果]A、B、C三组Ⅱ型胶原免疫组织化学检测阳性;MTT吸光度值和GAG含量检测结果均为:A组>C组>D组,B组>C组>D组,统计学有显著差异。[结论]兔BMSCs在一定条件下可以诱导成软骨细胞,TGF-β1和IGF-Ⅰ、bFGF在BMSCs向软骨细胞分化和增殖时起协同作用。[ Objective ] To study the methods of inducing bone mesenchymal stem cells （BMSCs） of rabbits into chondrocytes in vitro and the interaction of transforming growth factor β1 （ TGF - β1 ）, insulin - like growth factor - I （ IGF - I ） and basic fibroblast growth factor （bFGF） . [ Methods] BMSCs of rabbits were primarily cultured and subcuhured in vitro, and then divided into four groups according to the difference of factors： group A receiving TGF - β1 and bFGF; group B receiving TGF - β1 and IGF - I ; group C receiving TGF - β1 ; group D receiving no cell growth factor. After three weeks all the four groups were detected by methyl thiazolyl tetrazolium （MTT） assay, measurement of glycosaminoglycan （GAG） and immunohistochemistry. [Results] Immunohistochemical detection of collagen II was positive in groups A, B and C. The results of the MTT assay and the GAG content in groups A and B were obviously higher than those in groups C and D. [ Conclusion] Rabbit BMSCs can be induced into chondrocytes under certain conditions. TGF - β1, IGF - I and bFGF have synergy effect in the differentiation from BMSCs into chondrocytes.

关 键 词：骨髓间充质干细胞 软骨细胞 转化生长因子-Β1 胰岛素样生长因子-Ⅰ 碱性成纤维细胞生长因子 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]