摘要
目的 研究人胰岛素样生长因子 1 (hIGF 1 )基因转染关节软骨细胞获得稳定表达的可行性。方法 用脂质体转染法将含有hIGF 1cDNA的真核表达载体转染兔关节软骨细胞 ,经G4 1 8筛选 ,形成阳性细胞克隆。继续培养 4周 ,原位杂交检测hIGF 1的表达 ,免疫细胞化学检测Ⅱ型胶原的表达。结果 G4 1 8筛选后所获得的阳性细胞克隆 ,原位杂交检测表明hIGF 1基因得到稳定表达 ,免疫细胞化学检测显示转染后的软骨细胞仍表达Ⅱ型胶原。结论 外源性hIGF 1基因能够在软骨细胞内获得稳定表达 。
Objective To investigate the possibility of stable expression of human insulin like growth factor 1 (IGF 1) gene of cultured chondrocyte transfected with IGF 1 gene in vitro.Methods Eukaryotic expression vector pc DNA3.1 hIGF 1 plasmid containing hIGF 1 cDNA was transfected into rabbit articular chondrocyte with the help of profectamine. The stable expression of hIGF 1 in the positive cells by G418 selected was determined by in situ hybridization analysis and expression of type II collagen was detected by immunocytochemical analysis.Results Cell clones with G 418 resistance and positive signals of situ hybridization analysis of hIGF 1 were obtained. The type II collagen expression was stable by immunocytochemical analysis.Conclusion hIGF 1 gene can be transfected and stably expressed in articular chondrocyte. The articular chondrocyte transfected with hIGF 1 gene can still stably express type II collagen.
出处
《创伤外科杂志》
2003年第6期435-437,共3页
Journal of Traumatic Surgery
关键词
人胰岛素样生长因子1
软骨细胞
基因转染
表达
human insulin like growth factor 1
cartilaginous cell
gene transfection
expression
