兔骨髓间充质干细胞体外软骨向诱导的实验研究  

Experimental study of chondrogenic induction of rabbit bone marrow mesenchymal stem cells in vitro

作　　者：周静萍[1] 陶德韬[2] 施六霞[1] 

机构地区：[1]皖南医学院口腔系,安徽芜湖241002 [2]皖南医学院附属弋矶山医院口腔科,安徽芜湖241001

出　　处：《皖南医学院学报》2008年第1期9-11,15,共4页Journal of Wannan Medical College

基　　金：安徽省教育厅自然科学研究基金项目(2006KJ114C)

摘　　要：目的:体外分离培养兔骨髓间充质干细胞(MSCs),并定向诱导其向软骨细胞表型分化。方法:抽取兔股骨骨髓,经密度梯度离心与贴壁培养分离得到MSCs。用含有TGF-β、b-FGF、胰岛素、维生素C、转铁蛋白等的DMEM/Ham′s-F12培养基进行软骨向诱导。通过形态学观察,阿辛蓝-PAS特染及Ⅱ型胶原免疫组化染色,鉴定经过诱导后细胞的软骨细胞的表型。结果:兔MSCs原代细胞呈梭形,克隆样生长,经过诱导培养7d后,细胞形态由梭形、多角形变为椭圆形,胞浆丰富,细胞核和核仁清晰可见。阿辛蓝-PAS和Ⅱ型胶原染色阳性。结论:密度梯度离心与贴壁培养两种方法相结合可获得相对高纯度的MSCs,该细胞经诱导后可向软骨细胞分化。Objective： To isolate and culture the rabbit bone marrow mesenchyrnal stem cells （MSCs）for inducing them directly into phenotypic chondrocytes in vitro. Methods ; The marrow was extracted from femoral bone of New Zealand white rabbit for isolating MSCs by means of density gradient centrifugation and adherent culture. After cells induction by DMEM/Ham＇s-F12 medium containing TGF-β, b-FGF, insulin, VitC and transferrin, cell morphological observation, alcian bluePAS histochemistry and collagen type Ⅱ immuno-histochemistry were conducted for chondrogenic phenotype identification. Results ： Primary MSCs proliferated in visible colonies with spindle shape. After 7 days of induction, some of the MSCs developed from spindle-shaped or polygon to elliptic shape with abtmdant endochylema, clear nucleus and nucleoli, showing positive staining of alcian blue and collagen type Ⅱ. Conclusion ;Combination approach of density gradient centrifugation and adherent culture may generate highly purified MSCs, and the isolated MSCs can differentiate into cartilage cell under special culture condition.

关 键 词：骨髓间充质干细胞 软骨 细胞培养 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]