Induction of albumin expression in HepG2 cells using immobilized simplified recombinant fibronectin protein

Optimization of the extracellular environment is very important for hepatocyte function in vitro. We expressed new chimeric proteins of the collagen-binding domain (CBD) with cell attachment site (CAS) of fibronectin to enhance hepatocyte function, and the CBD-CAS proteins were immobilized on collag...

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Bibliographic Details
Published in:In vitro cellular & developmental biology. Animal 2013-06, Vol.49 (6), p.400-407
Main Authors: Nishida, Yuuki, Taniguchi, Akiyoshi
Format: Article
Language:English
Subjects:
Albumins
Albumins - biosynthesis
Albumins - metabolism
Animal Genetics and Genomics
Biomedical and Life Sciences
BIOTECHNOLOGY
Cell Biology
Cell Culture
Cell lines
Cell Movement - genetics
Collagen - chemistry
Collagen - genetics
Collagens
Complementary DNA
Cultured cells
Developmental Biology
Fibronectins - chemistry
Fibronectins - metabolism
Focal Adhesion Kinase 1 - metabolism
Focal adhesions
Gene Expression Regulation
Hep G2 Cells
Hepatocytes
Hepatocytes - metabolism
Humans
Integrins
Life Sciences
Messenger RNA
Phosphorylation
Protein Structure, Tertiary - genetics
Recombinant Fusion Proteins - chemistry
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Signal Transduction
Stem Cells
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Summary:Optimization of the extracellular environment is very important for hepatocyte function in vitro. We expressed new chimeric proteins of the collagen-binding domain (CBD) with cell attachment site (CAS) of fibronectin to enhance hepatocyte function, and the CBD-CAS proteins were immobilized on collagen-coated plates. We hypothesized that the high density of CAS would increase activity of the integrin-dependent intracellular signaling pathway, thus inducing hepatocyte function. Expression of albumin in the human hepatocyte cell line HepG2 was assessed on CBD-CAS-immobilized dishes. The results indicated that the CBD-CAS-immobilized plates induced albumin expression. Immobilized CBD-CAS induced activation of focal adhesion kinase and integrin-ligand clustering on the cell membrane. These results suggest that immobilized CBD-CAS improves the function of HepG2 cells. This system could therefore be applied to drug metabolism assay in the development of new drugs.
ISSN:1071-2690
1543-706X
DOI:10.1007/s11626-013-9594-4