In-depth physiological characterization of primary human hepatocytes in a 3D hollow-fiber bioreactor

被引:29
作者
Mueller, Daniel [1 ]
Tascher, Georg [1 ]
Mueller-Vieira, Ursula [2 ]
Knobeloch, Daniel [3 ]
Nuessler, Andreas K. [4 ]
Zeilinger, Katrin [5 ]
Heinzle, Elmar [1 ]
Noor, Fozia [1 ]
机构
[1] Univ Saarland, Biochem Engn Inst, D-66123 Saarbrucken, Germany
[2] Pharmacelsus GmbH, D-66123 Saarbrucken, Germany
[3] Charite, Dept Gen Visceral & Transplantat Surg, Campus Virchow Klinikum, D-13353 Berlin, Germany
[4] Tech Univ Munich, MRI, Dept Traumatol, D-81675 Munich, Germany
[5] Charite Campus Virchow Klinikum, Berlin Brandenburg Ctr Regenerat Therapies BCRT, Div Expt Surg, Bioreactor Grp, D-13353 Berlin, Germany
关键词
liver; metabolic activity; CYP; 450; cell metabolism; long-term studies; 3D cultivation; toxicity; HUMAN LIVER-CELLS; ONLINE GAS-ANALYSIS; STEM-CELLS; GENE-EXPRESSION; PHASE-I; SUPPORT; OXYGEN; DIFFERENTIATION; TOXICITY; COLLAGEN;
D O I
10.1002/term.418
中图分类号
Q813 [细胞工程];
学科分类号
摘要
As the major research focus is shifting to three-dimensional (3D) cultivation techniques, hollow-fiber bioreactors, allowing the formation of tissue-like structures, show immense potential as they permit controlled in vitro cultivation while supporting the in vivo environment. In this study we carried out a systematic and detailed physiological characterization of human liver cells in a 3D hollow-fiber bioreactor system continuously run for >2 weeks. Primary human hepatocytes were maintained viable and functional over the whole period of cultivation. Both general cellular functions, e. g. oxygen uptake, amino acid metabolism and substrate consumption, and liver-specific functions, such as drug-metabolizing capacities and the production of liver-specific metabolites were found to be stable for >2 weeks. As expected, donor-to-donor variability was observed in liver-specific functions, namely urea and albumin production. Moreover, we show the maintenance of primary human hepatocytes in serum-free conditions in this set-up. The stable basal cytochrome P450 activity 3 weeks after isolation of the cells demonstrates the potential of such a system for pharmacological applications. Liver cells in the presented 3D bioreactor system could eventually be used not only for long-term metabolic and toxicity studies but also for chronic repeated dose toxicity assessment. Copyright (C) 2011 John Wiley & Sons, Ltd.
引用
收藏
页码:E207 / E218
页数:12
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