hIL-10基因修饰的L02肝细胞的克隆培养和最高表达株的筛选

目的:确认hIL-10基因修饰的L02肝细胞的克隆培养可实现hIL-10在L02肝细胞中的高效表达．方法:通过构建真核质粒表达载体pchIL-10, 并纯化后转染L02肝细胞．通过G418的压力选择获得hIL-10高表达的克隆株,并以ELISA测定其表达水平．结果:经过测序和酶切验证,真核质粒表达载体pchIL-10构建成功．电泳显示一长约540 bp 条带．hIL-10基因转导可在L02肝细胞中实现 hIL-10的高效表达．最高表达株表达量为每小时69．875 ng/106细胞．结论:hIL-10基因修饰的L02肝细胞的克隆培养可实现hIL-10的高效表达,为抗肝纤维化、肝硬化提供有效途径．

Clone culture of human interleukin-10 gene modified L02 hepatocytes and selection of cell strain with most interleukin-10 expression

AIM: To clone and culture human interleukin-10 (IL-10) gene modified L02 hepatocytes and select the cell strain highly expressing IL-10. METHODS: With preparation of previously constructed and purified eukaryotic expression plasmid vector pchIL-10, L02 hepatocytes were transfected and then the positive clones were selected with the help of G418 pressure. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of human IL-10 in the cells strain. RESULTS: Sequencing and restriction endo-nuclease digestion confirmed that eukaryotic expression plasmid vector pchIL-10 was constructed successfully, and electrophoresis show a band of 540 bp. hIL-10 gene was highly expressed in L02 hepatocytes and the highest expression level was 69.875 ng/106 cell per hour. CONCLUSION: Human interleukin-10 (IL-10) gene modified L02 hepatocytes can highly expresses hIl-10, which may be used in the antifibrotic or cirrhotic treatment.