|Cancer||Cell death||Cell cycle||Cytoskeleton||Exo/endocytosis||Differentiation||Division||Organelles||Signalling||Stem cells||Trafficking|
Cell Biology International (2010) 34, S38 (Printed in Great Britain)
Establishment and characterization of a liver stem cell line with stable expression of functional ALB promoter and luciferase reporter gene
Yun He12, Yang Bi12, Weiyu Zhang1 and Tongchuan He12
1Stem Cell Biology and Therapy Laboratory, Children's Hospital of Chongqing Medical University, Chongqing 400014, China, and 2Molecular Oncology Laboratory, The University of Chicago Medical Center, Chicago, IL 60637, USA
Albumin (ALB) has been widely used as a liver-specific marker for the detection of hepatocyte differentiation. In this report, we constructed a liver stem cell line with stable expression of functional ALB promoter and luciferase reporter gene. ALB promoter was amplified by PCR, then ligated with pBGLuc plasmid containing luciferase reporter gene to get a pBGLuc-ALB plasmid. Relative ALB-GLuc activity of pBGLuc-ALB transfected HEK293, HP14.5 (Hepatic progenitor cells derived from E14.5 mouse fetal liver), LC14 (Live Cells derived from 14-day old mouse liver) and Hepa1-6 cells was coincident with ALB expression level in each cell lines. Afterward, HP14.5 cells were infected by ALB-GLuc retrovirus to get stable cell line, which could survive in medium containing high concentration of Blasticidin. Stable cell line were cultured with dexamethasone and HGF induction medium in vitro, relatively ALB-Gluc activity gradually increased from 3 to 12 days after induction and had a uniform increasing tendency compared with ALB expression detected by immunofluorescence. Therefore, we successfully constructed a liver stem cell line with stable expression of ALB promoter and luciferase reporter gene for further study of hepatocyte differentiation.
Published online 1 August 2010, doi:10.1042/CBI034S038b
© The Author(s) Journal compilation © 2010 Portland Press Limited