title: In-depth physiological characterization of primary human hepatocytes in a 3D hollow-fiber bioreactor.
abstract: 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.
authors: Biochemical Engineering Institute, Saarland University, Saarbruecken, Germany.
authors: Daniel
authors: D
authors: Mueller
authors: Georg
authors: G
authors: Tascher
authors: Ursula
authors: U
authors: Müller-Vieira
authors: Daniel
authors: D
authors: Knobeloch
authors: Andreas K
authors: AK
authors: Nuessler
authors: Katrin
authors: K
authors: Zeilinger
authors: Elmar
authors: E
authors: Heinzle
authors: Fozia
authors: F
authors: Noor
mesh_terms: D001219:Aspartate Aminotransferases / Q000378:metabolism; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology*; D006801:Humans; D009928:Organ Specificity; D010101:Oxygen Consumption; D004364:Pharmaceutical Preparations / Q000378:metabolism; D013379:Substrate Specificity; D013997:Time Factors
chemical_list: D004364:Pharmaceutical Preparations; D003577:Cytochrome P-450 Enzyme System; D001219:Aspartate Aminotransferases