体外诱导大鼠脂肪源间充质干细胞成肌腱潜能的研究  

Inducing rat adipose derived mesenchymal stem cells according to tendon potential in vitro

作　　者：侯凯[1] 李梅[1] 李金茹[1] 杨逸昆[1] 李纯[1] 王文秀[1] 黄德清[2] 

机构地区：[1]天津医科大学基础医学院微生物学教研室,天津300070 [2]天津市第三医院骨科,天津300250

出　　处：《中国现代医学杂志》2011年第8期929-933,共5页China Journal of Modern Medicine

基　　金：2008年天津市应用基础及前沿技术研究计划项目(No:08JCYBJC27100);2007年天津市卫生局科技基金项目(No:07KR01)

摘　　要：目的体外诱导脂肪源间充质干细胞(adipose tissue-derived mesenchymal stem cells,AT-MSCs)成肌腱细胞,为临床肌腱损伤的修复提供理论基础。方法从SD大鼠腹腔脂肪组织中分离出脂肪源间充质干细胞,经生长分化因子-5(growth differentiation factor 5,GDF-5)诱导,使其向肌腱细胞分化。应用形态观察和RT-PCR等方法,从形态和分子水平上对诱导分化的细胞进行鉴定。结果从SD大鼠腹腔脂肪组织中分离出的AT-MSCs呈散在分布,排列不规则,以星形和多角形细胞为主,能稳定增殖传代。经5、10 ng/mL GDF-5诱导8 d后,大部分细胞呈梭形。RT-PCR结果显示经GDF-5诱导的细胞,Ⅰ型和Ⅲ型胶原蛋白的表达明显高于未经诱导的对照组;GDF-5诱导的细胞Bcl-2/Bax均高于对照组。结论从脂肪组织中可以获得具有多向分化能力的AT-MSCs,并能在体外稳定传代。经GDF-5诱导后,AT-MSCs有向肌腱细胞分化的倾向。GDF-5在一定时间内对细胞凋亡有抑制作用,有利于AT-MSCs向肌腱细胞的分化。AT-MSCs有可能替代骨髓间充质干细胞作为组织工程学的种子细胞,用于肌腱损伤的修复。【Objective】 To verify that rat adipose tissue-derived mesenchymal stem cells（AT-MSCs） can differentiate into tenocytes in vitro,so as to lay a foundation for the cell-based transplantation therapy of tendon injury.【Methods】 AT-MSCs were isolated from abdominal adipose tissue of adult Sprague Dawley rats and induced towards tenocytes in the medium containing growth differentiation factor 5（GDF-5）.The appearance of the differentiated cells was observed.Meanwhile,the expression level of type I,II,III collagens and Bcl-2,Bax（Bcl-2 associated X protein） were detected to evaluate the process of differentiation by means of semi-quantitative reverse transcription polymerase chain reaction（RT-PCR）.【Results】 Rat AT-MSCs presented diffused distribution and proliferated stably in vitro.The appearance of the differentiated AT-MSCs towards tenocytes varied from the irregular-arranged multangular cells into parallel-arranged,spindle shape or fusiform cells.The results of RT-PCR show the gene expressions of type I and III collagens,which were characteristics of tendon cells,were obviously upgraded in the induced cells by GDF-5,meanwhile,expression ratio of Bcl-2 and Bax（Bcl-2/Bax） was obviously higher in the induced cells by GDF-5 than that of the control.【Conclusion】 Rat AT-MSCs from abdominal adipose tissue can be passaged stably and differentiated into tendon cells in vitro.GDF-5 can inhibit AT-MSCs apoptosis within a certain period of time.It suggests that AT-MSCs could become promising seed cells of tissue engineering during the therapy of tendon repair instead of marrow stromal cells.

关 键 词：脂肪源间充质干细胞 肌腱细胞 生长分化因子-5 细胞凋亡 

分 类 号：R-332[医药卫生]