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
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代文杰, 胡 震, 武林枫, 姜洪池, 吴耀华,
白细胞介素10;基因转导;肝纤维化;肝硬化,代文杰,胡震,武林枫,姜洪池,吴耀华,潘尚哈,代文杰,通讯作者,Clonecultureof
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代文杰, 胡震, 武林枫, 姜洪池, 吴耀华, 潘尚哈, 哈尔滨医科大学第一临床医学院肝胆胰外科, 黑龙江省肝脾外科重点实验室 黑龙江省哈尔滨市 150001
代文杰, 博士, 博士后, 主要从事肝胆胰外科、器官移植、外科分子细胞生物学研究.
国家自然科学基金资助项目, No. 30070739, 30300340
黑龙江省自然科学基金资助项目, No. D01-04
通讯作者: 代文杰, 150001, 黑龙江省哈尔滨市, 哈尔滨医科大学第一临床医学院肝胆胰外科, 黑龙江省肝脾外科重点实验室. wenjdai@yahoo.com.cn
电话: 0451-53600281
收稿日期: 2006-03-30 接受日期: 2006-04-13
Clone culture of human interleukin-10 gene modified L02 hepatocytes and selection of cell strain with most interleukin-10 expression
Wen-Jie Dai, Zhen Hu, Lin-Feng Wu, Hong-Chi Jiang, Yao-Hua Wu, Shang-Ha Pan
Wen-Jie Dai, Zhen Hu, Lin-Feng Wu, Hong-Chi Jiang, Yao-Hua Wu, Shang-Ha Pan, Department of Hepatobiliary and Pancreatic Surgery, the First Clinical Medical College of Harbin Medical University; Key Laboratory of Hepatosplenic Surgery, Heilongjiang Province, Harbin 150001, China
Supported by National Natural Science Foundation of China, No. 30070739 and 30300340, and the Natural Science Foundation of Heilongjiang Province, No. D01-04
Correspondence to: Wen-Jin Dai, Department of General Surgery, the First Clinical Medical College of Harbin Medical University; Key Laboratory of Hepatosplenic Surgery, Heilongjiang Province, Harbin 150001, China. wenjdai@yahoo.com.cn
Received: 2006-03-30 Accepted: 2006-04-13
Abstract
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 endonuclease digestion confirmed that eukaryotic expression plasmid vector pchIL-10 was constructed successfully, and electrophoresis show a band of 540 bp ......
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