Wolff J A, Yee J K, Skelly H F, Moores J C, Respess J G, Friedmann T, Leffert H
Proc Natl Acad Sci U S A. 1987 May;84(10):3344-8. doi: 10.1073/pnas.84.10.3344.
Differentiated primary rat hepatocyte cultures have been infected with retroviral vectors expressing human hypoxanthine/guanine phosphoribosyltransferase or the transposon Tn5 neomycin-resistance gene. Expression of the markers was detected only after infection of the cells during a short period of cell replication and transient dedifferentiation from days 1 to 5 of culture. Provirus integrated during that period remains fully expressed during the entire subsequent stationary period of culture up to at least 25 days. Selection with the neomycin analogue G418 of cells infected with the neomycin vector led to the appearance of cells with hepatocyte morphology in which newly synthesized albumin was detectable by immunoprecipitation, indicating successful infection of hepatocytes.
已用表达人次黄嘌呤/鸟嘌呤磷酸核糖转移酶或转座子Tn5新霉素抗性基因的逆转录病毒载体感染分化的原代大鼠肝细胞培养物。仅在培养第1至5天的短时间细胞复制和短暂去分化期间感染细胞后,才检测到标志物的表达。在此期间整合的前病毒在随后至少25天的整个静止培养期内仍能充分表达。用新霉素载体感染的细胞经新霉素类似物G418筛选后,出现了具有肝细胞形态的细胞,通过免疫沉淀可检测到新合成的白蛋白,这表明肝细胞感染成功。