TGF-β1基因修饰诱导脂肪干细胞成软骨分化的实验研究  

Adipose tissue derived stem cells transferred with TGF-β_1 gene and differentiated into chondrocytes

作　　者：王洪林[1] 闵繁红 王岩峰[2] 谢永[1] 张金刚[1] 栾洪佳 侯旭东[1] 

机构地区：[1]黑龙江省佳木斯市中心医院骨外科,154002 [2]中国医科大学附属第一医院骨科,辽宁省沈阳市110001

出　　处：《中国矫形外科杂志》2008年第17期1347-1350,共4页Orthopedic Journal of China

摘　　要：[目的]本研究通过选择对软骨细胞ECM合成具有促进作用的TGF-β1转染脂肪干细胞(ADSCs),观察转染后目的基因的稳定表达和对软骨细胞外基质(ECM)的两种重要组分:Ⅱ型胶原(typeⅡcollage,ColⅡ)和ag-grecan的合成,为构建新型的组织工程软骨提供实验依据。[方法]TGF-β1基因转染脂肪干细胞及阳性细胞克隆株的筛选;RT-PCR检测TGF-β1、纤连蛋白(fibronectin,FN)、ColⅡ、Aggrecan的表达;Western-blot检测TGF-β1。[结果]RT-PCR结果显示:实验组(TGF-β1稳定转染组)的TGF-β1、FN、ColⅡ、Aggrecan的表达较空染组和正常对照组均明显增多(P<0.01);Western blot检测实验组(TGF-β1稳定转染组)TGF-β1蛋白的表达较空染组和正常对照组均明显增多(P<0.01)。[结论]成功分离培养脂肪干细胞;以脂质体介导法将TGF-β1目的基因成功转染ADSCs,转染后成软骨分化的特异性细胞外基质增多,表明转染TGF-β1基因可以促使ADSCs向软骨细胞方向分化,从而为软骨组织工程提供新的思路和方法。[ Objective] To induce TGF - β1 gene which can increase ECM synthesis into adipose tissue derived stem cells （ADSCs） by employing gene transfer techniques and observe whether TGF - β1 gene could expresse continuously and whether type Ⅱ collagen and aggrecan are synthesized in order to provide experimental data for constructing tissue - engineering cartilage. [Method] ADSCs Were transferred with TGF- β1 gene , TGF- β1, FN, Col Ⅱ and aggrecan were detected with RT- PCR and TGF - β1 protein was detected with Western - blot. [ Result] RT - PCR demonstrated that the expression of TGF - β1, FN, Col Ⅱ and aggrecan in the TGF - β1 gene transferred groups increased more evidently than non - gene groups and control groups （P 〈 0. 01 ） . Western blot demonstrated that TGF - β1 protein in the TGF - β1 gene transferred groups in- creased more evidently than non - gene groups and control groups significantly （P 〈 0. 01 ） . [ Conclusion] ADSCs of Wistar rats could be separated and cultured successfully. After TGF - β1 gene was transferred stably into ADSCs, the specific extracellular matrix of chondrocyte differentiation increased evidently. The results demonstrated that TGF - β1 gene could induce ADSCs into chondrocytes. The experiment provides a new methed for constructing tissue - engineering cartilage.

关 键 词：转化生长因子β1(TGF-β1) 脂肪干细胞(ADSCs) 基因转染 RT—PCR Western—blot 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]