RRC ID 4770
Author Kobayashi N, Noguchi H, Westerman KA, Watanabe T, Matsumura T, Totsugawa T, Fujiwara T, Leboulch P, Tanaka N.
Title Cre/loxP-based reversible immortalization of human hepatocytes.
Journal Cell Transplant
Abstract An ideal alternative to the primary human hepatocytes for hepatocyte transplantation would be to use a clonal cell line that grows economically in culture and exhibits the characteristics of differentiated, nontransformed hepatocytes following transplantation. The purpose of the present studies was to establish a reversibly immortalized human hepatocyte cell line. Human hepatocytes were immortalized with a retroviral vector SSR#69 expressing simian virus 40 large T antigen (SV40Tag) gene flanked by a pair of loxP recombination targets. One of the resulting clones, NKNT-3, showed morphological characteristics of liver parenchymal cells and expressed the genes of differentiated liver functions. NKNT-3 cells offered unlimited availability. After an adenoviral delivery of Cre recombinase and subsequent differential selection, efficient removal of SV40Tag from NKNT-3 cells was performed. Here we represent that elimination of the retrovirally transferred SV40Tag gene can be excised by adenovirus-mediated site-specific recombination.
Volume 10(4-5)
Pages 383-6
Published 2001
PMID 11549058
MeSH Cell Line* Cell Transplantation Culture Media, Serum-Free Gene Transfer Techniques* Genetic Vectors Hepatocytes / physiology* Hepatocytes / transplantation Humans Integrases / metabolism* Liver / physiology Recombination, Genetic Retroviridae / genetics Retroviridae / metabolism Transduction, Genetic Viral Proteins / metabolism*
IF 2.885
Times Cited 15
WOS Category TRANSPLANTATION MEDICINE, RESEARCH & EXPERIMENTAL CELL & TISSUE ENGINEERING
Resource
DNA material AxCALacZ (RDB1745)