软骨形态发生蛋白1诱导犬真皮成纤维细胞向软骨细胞表型分化的实验研究

Study of CDMP1 Inducing DFs into Chondrogenic Lineage in Vitro

目的探讨软骨形态发生蛋白1(Cartilage-derived morphogenetic protein-1,CDMP1)诱导体外培养的犬真皮成纤维细胞(Dermal fibroblasts,DFs)向软骨细胞表型分化的可行性。方法应用CDMP1细胞诱导液对体外扩增至第4代的犬DFs进行诱导培养,观察细胞的形态变化,以免疫荧光染色和RT-PCR检测细胞中软骨特异蛋白的表达情况。结果诱导培养6 d后,实验组部分DFs呈星形或多角形,免疫组化染色显示有软骨特异的II型胶原分泌,RT-PCR检测也表明有Ⅱ型胶原、aggrecan和SOX9表达。但是诱导培养10 d后,均转为阴性。结论 CDMP1可以诱导犬DFs向软骨细胞表型分化,有可能成为组织工程化纤维软骨的种子细胞。

Objective To explore the feasibility of cartilage-derived morphogenetic protein-1（CDMP1） inducing dermal fibroblasts（DFs） into chondrogenic lineage.Methods The fourth passage of DFs isolated from canine were induced by CDMP1.The morphological changes were observed and the expression of cartilage specific proteins were detected by immunofluorescent staining and RT-PCR.Results After inducing 6 days,stellar-like or polygonal-like cells were observed,and cartilage specific collagen type Ⅱ was detected by immunofluorescent staining as well as RT-PCR examination.Other cartilage specific proteins as aggrecan and SOX9 were detected by RT-PCR.But these effects could not be maintained after inducing 6 days.Conclusion CDMP1 could induce DFs into chondrogenic lineage,and CDMP1 induced DFs is promising to be the seed cell of engineered fibrochondrocytes.