过表达SOX9改良软骨组织工程种子细胞的可行性研究  

Feasibility Study on Overexpressing SOX9 Modified Cartilage Tissue Engineering Seed Cells

作　　者：胡澜 祝子涵 吴俊楠 徐瑶 肖建华[1] HU Lan;ZHU Zi-han;WU Jun-nan;XU Yao;XIAO Jian-hua(College of Veterinary Medicine,Northeast Agricultural University,150030)

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《动物医学进展》2020年第12期68-73,共6页Progress In Veterinary Medicine

摘　　要：自体软骨细胞是软骨组织工程最佳种子细胞,但存在细胞外基质合成能力的局限性。为了获得合成细胞外基质能力更强的种子细胞,构建了pcDNA3.1(+)-SOX9哺乳动物过表达载体质粒,脂质体转染分离培养的大鼠关节软骨细胞。免疫荧光法和Western blot检测SOX9过表达效率,实时荧光定量PCR和Western blot检测CollagenⅡ(Ⅱ型胶原)和Aggrecan(蛋白聚糖)mRNA和蛋白的表达,阿利新蓝染色检测细胞外基质合成情况。结果表明,成功过表达软骨细胞的SOX9基因,细胞外基质分泌能力明显增强,过表达SOX9基因能够促进细胞外基质分泌,通过转染过表达SOX9来改良软骨种子细胞具有可行性。Autologous chondrocyte is the best cell resource for cartilage tissue engineering which is limited in the function of synthesize extracellular matrix.Recombinant pcDNA3.1(+)-SOX9 overexpression vector was constructed and transfected into rat articular cartilage cell with liposome to obtain seed cells with stronger ability of extracellular matrix synthesis.The overexpression efficiency of SOX9 was detected by immunofluorescence and Western blot.The mRNA and protein levels for CollagenⅡand Aggrecan were quantified by real-time quantitative polymerase chain reaction and Western blot.Extracellular matrix was assessed by straining of Alcian blue.The experimental result showed that SOX9 gene was overexpressed successfully and the ability of synthesizing extracellular matrix was significantly enhanced.The results illustrated that overexpression of SOX9 gene promotes the synthesis of extracellular matrix,and is a feasible way to ameliorate articular chondrocytes.

关 键 词：SOX9基因 pcDNA3.1(+)哺乳动物过表达载体 种子细胞 细胞外基质 

分 类 号：S854.56[农业科学—临床兽医学] S858.91[农业科学—兽医学]