永生化人胎肝细胞系LC-1的建立

目的 建立人源性永生化肝细胞.方法 利用含有SV40T和EGFP基因的双顺反子逆转录病毒表达载体pLNCTIGlox,转染包装细胞PT67;取病毒上清液感染原代培养的人胎肝细胞,用G418筛选成功转导的永生化细胞系;用细胞计数法测定永生化细胞增殖特性,免疫细胞化学染色检测肝细胞特征蛋白表达,放免法和自动生化仪检测清蛋白和尿素氮合成功能.结果 获得的一个细胞系命名为LC-1,其细胞群体倍增时间约为17h,可表达清蛋白、CK18肝细胞特征和CK19等胆管特征蛋白,具有清蛋白与尿素氮合成功能.结论 成功建立了具有良好的增殖能力、分化状态、肝细胞生物学特征的永生化肝细胞系.

Establishment of immortalized human fetal hepatocyte line LC-1

Objective To establish immortalized human hepatocyte lines.Methods PT67 cells were transfected with pLNCTIGlox,an ambicistron retroviral expression vector previously constructed by our study group and containing ST40 and EGFP.Primary human fetal hepatocytes were infected with virus-contained supertanants and the successfully transducted cell lines were screened with G418.The proliferation characteristics of the cells were evaluated with cytometry.Hepatocyte-specific protein expression was detected with immu...