小鼠胚胎瘤来源细胞系ATDC5的体外软骨分化诱导研究  

The model of chondrogenic differentiation in vitro induced by mouse embryonal carcinoma-derived cell line ATDC5

作　　者：金旻[1] 戚华兵[1] 苏楠[1] 杜晓兰[1] 杨京[1] 赵玲[1] 陈林[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所全军战刨伤中心,创伤、烧伤与复合伤国家重点实验室,重庆400042

出　　处：《中国骨质疏松杂志》2009年第10期737-740,共4页Chinese Journal of Osteoporosis

基　　金：国家自然科学基金重点资助项目(30530410);国家杰出青年科学基金资助项目(30425023)

摘　　要：目的在体外建立稳定有效的软骨细胞分化模型。方法复苏ATDC5细胞,在倒置显微镜下观察细胞形态及其生长情况,细胞90%融合时分别用不含Vc和含Vc的诱导培养基培养进行分化诱导。诱导21 d,阿力新蓝染色,RT-PCR检测Ⅱ、Ⅹ型胶原表达进行鉴定。结果含50μg/mL Vc的诱导培养基诱导1周便可见明显的软骨小结,细胞产生软骨基质明显增多,Ⅱ及Ⅹ型胶原表达明显增高,且Ⅹ型胶原表达呈提前。结论利用ATDC5细胞系可成功建立软骨细胞分化体外模型。Objective To set up a stable and effective model in vitro of chondrogenic differentiation. Methods ATDC5 cells were reanimated from liquid nitrogen, and cultured in the maintenance medium consisting of a 1 ： 1 mixture of DME and Ham＇s F-12 medium containing 5% FBS. The morphological changes and growth feature were observed under the inverted microscope each day. When ATDC5 cells rapidly proliferated to form 90 % confluency, the cells were cultured in differentiation medium supplemented with Vc or no Vc for 21 d. The extent of chondrogenie differentiation was examined by aleian blue staining and RT-PCR detection of the expression level of mRNA of type Ⅱ collagen and type X collagen. Results For ATDC5 cells cultured in differentiation medium containing 50 μg/ml Vc, cartilaginous nodules were detectable apparently at 7 d after chondregenic differentiation induction, the synthesis of glycosaminoglycans （GAGs） were increased, and the expression levels of type Ⅱcollagen and type X collagen mRNA were also upregulated, with earlier expression of type X collagen. Conclusion The method used in this work can provide an in vitro chondrogenic differentiation model.

关 键 词：ATDC5 软骨细胞 分化 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]