Growth and differentiation of colony-forming human hepatocytes in vitro

Journal of Hepatology Volume 44 Issue 4 Page 749-757 published_at 2006-04
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Title ( eng )
Growth and differentiation of colony-forming human hepatocytes in vitro
Creator
Yamasaki Chihiro
Tateno Chise
Aratani Akio
Ohnishi Chimoto
Katayama Shigeru
Kohashi Toshihiko
Hino Hiroshi
Marusawa Hiroyuki
Source Title
Journal of Hepatology
Volume 44
Issue 4
Start Page 749
End Page 757
Abstract
Background/Aims: Parenchymal hepatocytes (PHs) of rat contain colony-forming parenchymal hepatocytes (CF-PHs) as a small fraction. We aimed to demonstrate the presence of CF-PHs in humans and characterize them with respect to growth and differentiation potential. Methods: Human PHs were co-cultured with Swiss 3T3 cells in the medium containing human serum, EGF, nicontinamide, and ascorbic acid 2- phosphate. To examine differentiation potential hepatocytes were cultured on gels of Matrigel Matrix. Results: Few PHs formed colonies, the colony-forming efficiency being as low as 0.01-0.09%.0 The CF-PHs could be subcultured up to 7 passages. They showed a liver epithelial cell-like morphology, and immunocytochemically positive for albumin (ALB), cytokeratin (CK) 7, 8, 18, and 19 in a pre- and early phase-confluence, whereas they showed a typical differentiated hepatocyte-like morphology, and positive for α1-antitrypsin, but negative for CK7 and 19 in condensed regions at confluence. The CF-PHs at late confluence expressed mRNAs of ALB, HNF4, and isoforms of cytochrome P450 at low levels. However, when cultured on Matrigel, these cells expressed them at high levels comparable to those of original PHs. Conclusions: We concluded that the human liver contains highly replicative hepatic progenitor-like cells as a minute population that retain a normal differentiation potential.
Keywords
Progenitor cells
Human hepatocytes
Spheroids
Matrigel
Hepatocyte propagation
Albumin
HNF4
Cytochrome P450
Cytokeratins
3-dimensional culture
NDC
Medical sciences [ 490 ]
Biology [ 460 ]
Language
eng
Resource Type journal article
Publisher
Elsevier
Date of Issued 2006-04
Rights
Copyright (c) 2006 Elsevier Ltd.
Publish Type Author’s Original
Access Rights open access
Source Identifier
[ISSN] 0168-8278
[DOI] 10.1016/j.jhep.2005.10.028
[NCID] AA10459105
[PMID] 16469405
[DOI] http://dx.doi.org/10.1016/j.jhep.2005.10.028 isVersionOf