CDMP1诱导成纤维细胞向成软骨细胞表型分化的实验研究  

Experimental study of chondrogenic differentiation of fibroblast induced with CDMP1

作　　者：尹烁[1] 崔磊[1] 李纲[1] 刘伟[1] 曹谊林[1] 

机构地区：[1]上海第二医科大学第九人民医院整复外科上海市组织工程研究与开发中心,上海200235

出　　处：《上海第二医科大学学报》2004年第4期254-257,共4页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：国家重点基础研究发展规划资助项目("973"项目)(1999054300);上海市青年科技启明星计划资助(OOQD145025).

摘　　要：目的 应用软骨形态发生蛋白-1(CDMP1)生长因子,体外诱导真皮成纤维细胞向成软骨细胞表型分化,探讨其作为组织工程化软骨种子细胞的可行性。方法 取成人真皮成纤维细胞,体外扩增至第2代,以加入CDMP1生长因子(100 ng/mL)的含10％胎牛血清的F-12培养液诱导,每3 d换液1次,进行单层培养,对照组为不加CDMP1培养的成纤维细胞。7d后,免疫荧光检测Ⅱ型胶原分泌,RT-PCR检测Aggrecan、Ⅱ型胶原mRNA表达。结果 诱导后成纤维细胞细胞形态由梭形向软骨细胞样多角形、多边形转变。Ⅱ型胶原免疫荧光检测表达阳性,RT-PCR检测Aggrecan、Ⅱ型胶原mRNA表达阳性。结论 成纤维细胞在CDMP1生长因子诱导下能向成软骨细胞表型分化,并能分泌软骨细胞特异性基质,有可能成为软骨组织工程新的种子细胞来源。Objective To investigate the feasibility of inducing chondrogenic differentiation of dermal fibroblasts with cartilage-derived morphogenetic protein-1 ( CDMP1) and the potential of induced cells as seeding cells in cartilage tissue engineerings. Methods Human dermal fibroblasts of passage 2 were induced with CDMP-1 (100 ng/ ml) in F-12 medium containing 10% FBS. Culture medium was changed every 3 days in monolayer culture, non-induced fibroblast was used as control. After 7 days, immunofluorescence was used to detecte the secretion of type Ⅱ collagen, RT-PCR was used to detect mRNA expression of aggrecan and type Ⅱ collagen. Results After induction , fibroblasts changed their morphology from spindle shape to typical polygon shape of chondrocytes. Type Ⅱ collagen expression was detected by immunofluorescence, mRNA expression of Aggrecan and collagen type Ⅱ were detected by RT-PCR. Conclusion Fibroblasts can be induced to express chondrogenic phenotype with CDMP1 and produce specific matrix of chondrocytes.

关 键 词：组织工程 软骨细胞 成纤维细胞 软骨形态发生蛋白-1 胶原 免疫荧光法 细胞诱变 

分 类 号：Q813.5[生物学—生物工程] R318.17[医药卫生—生物医学工程]