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Journal of Biochemistry Advance Access published online on January 27, 2009
Journal of Biochemistry, doi:10.1093/jb/mvp019
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© The authors 2009. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

Maturation of the extracellular matrix and cell adhesion molecules in layered co-cultures of HepG2 and endothelial cells

Maki Ohno1,2, Kiyoto Motojima2, Teruo Okano3 and Akiyoshi Taniguchi1

1Advanced Medical Materials Group, Biomaterials Center, National Institute for Materials Science, 1-1, Namiki, Tsukuba, Ibaraki 305-0044, Japan
2Department of Biochemistry, Meiji Pharmaceutical University, 2-522-1, Noshio, Kiyose, Tokyo 204-8588, Japan
3Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, 8-1, Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan

To whom correspondence should be addressed: Akiyoshi Taniguchi, Ph.D., Cell-Sensing Group, Biomaterials Center, National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan, Phone: +81-29-860-4505; Fax: +81-29-860-4714, E-mail: TANIGUCHI.Akiyoshi{at}nims.go.jp

Received December 14, 2008; Accepted January 15, 2009


  Abstract

We previously reported that using thermo-responsive culture surfaces, a layered co-culture was achieved by placing an endothelial cell sheet onto a layer of human hepatoma cell line HepG2 in order to develop a culture model that mimics hepatic lobules. In the layered co-culture cells, the expression levels of liver-specific genes gradually increased. A cross-sectional view of the layered co-culture cells showed that the thickness of the layer slowly increased after layering, as did extracellular matrix (ECM) deposition around HepG2 cells. In this study, we report that the molecular compositions of ECM and cell adhesion molecules changed in the layered co-culture cells. Gene expression of integrin {alpha}4 and decorin gradually increased after layering, and the time-course pattern of these genes was correlated with that of liver-specific genes. Moreover, the layered co-culture system has the ability to assemble a branching network of fibronectin fibrils. These results suggest that a vastly different extracellular environment in layered co-culture cells may induce an increase in liver-specific functions.

Key Words: Hepatocyte, co-culture, ECM, Gene expression, cell sheet technology


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