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Construction of three‐dimensional liver tissue models by cell accumulation technique and maintaining their metabolic functions for long‐term culture without medium change
Author(s) -
Matsuzawa Atsushi,
Matsusaki Michiya,
Akashi Mitsuru
Publication year - 2015
Publication title -
journal of biomedical materials research part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.849
H-Index - 150
eISSN - 1552-4965
pISSN - 1549-3296
DOI - 10.1002/jbm.a.35292
Subject(s) - materials science , term (time) , metabolic activity , biomedical engineering , biological system , cell culture , tissue culture , liver tissue , biochemical engineering , biochemistry , biology , engineering , endocrinology , in vitro , genetics , physics , quantum mechanics
Three‐dimensional (3D) hepatocyte cultures have attracted much attention to obtain high biological functions of hepatocyte for pharmaceutical drug assessment. However, maintaining the high functions for over one month is still a key challenge although many approaches have been reported. In this study, we demonstrate for the first time simple and rapid construction of 3D‐hepatocyte constructs by our cell accumulation technique and their high biological functions for one month, without any medium change. The human hepatocyte carcinoma (HepG2) cells were coated with ∼7 nm‐sized extracellular matrix (ECM) films consisting of fibronectin (FN) and gelatin (G), and then incubated in cell culture insert to construct 3D‐tissue constructs for 24 h. The thickness of obtained 3D‐HepG2 constructs was easily controlled by altering seeding cell number and the maximum is over 100 μm. When a large volume of culture media was employed, the 3D‐constructs showed higher mRNA expression of albumin and some cytochrome P450 (CYP) enzymes as compared to general two‐dimensional (2D) culture. Surprisingly, their high cell viabilities (over 80%) and high mRNA expressions were successfully maintained without medium change for at least 27 days. These results demonstrate novel easy and rapid technique to construct 3D‐human liver tissue models which can maintain their high functions and viability for 1 month without medium change. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 1554–1564, 2015.