Expanding hepatocytes in vitro before cell transplantation: donor age-dependent proliferative capacity of cultured human hepatocytes

Scandinavian Journal of Gastroenterology
J WalldorfW E Fleig

Abstract

For hepatocyte transplantation as well as experimental purposes, it would be advantageous to be able to expand human hepatocytes in vitro. However, under serum-free conditions, even with supplements of HGF (hepatic growth factor) and EGF (epidermal growth factor), proliferation of human hepatocytes is hampered. The aim of this study was to identify differences in the proliferative capacity of cultured primary human hepatocytes related to the age of the liver donors. Proliferation was determined by BrdU-uptake, ploidy was measured using propidium iodide staining and flow cytometry, and the expression of cell cycle related proteins was determined by Western blotting. During the initial culture, juvenile hepatocytes proliferated better than adult hepatocytes. The proliferation rate declined to barely detectable levels after 8 days in culture in both juvenile and adult hepatocytes. The higher proliferative capacity of juvenile hepatocytes was associated with a larger fraction of diploid cells and a higher viability. The expression of regulatory cell cycle related proteins was higher in juvenile than in adult hepatocytes. The proliferation of human hepatocytes in vitro is critically related to a large fraction of diploid hepatocytes...Continue Reading

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Citations

Sep 18, 2007·Tissue Engineering·Kirstin LinkeHeike Mertsching
Nov 28, 2015·Expert Opinion on Drug Metabolism & Toxicology·Zdenek Dvorak
Mar 16, 2013·Liver International : Official Journal of the International Association for the Study of the Liver·Anwar A PalakkanJames A Ross
Jan 27, 2015·Journal of Hepatology·Maria Paola SerraEzio Laconi
Feb 3, 2007·Cell Transplantation·Trinidad SerranoRobin D Hughes

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