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

J Tissue Eng Regen Med. 2011 Aug;5(8):e207-18. doi: 10.1002/term.418. Epub 2011 Mar 27.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aspartate Aminotransferases / metabolism
  • Bioreactors*
  • Cell Culture Techniques / instrumentation*
  • Cell Culture Techniques / methods*
  • Cell Survival
  • Cells, Cultured
  • Cytochrome P-450 Enzyme System / metabolism
  • Hepatocytes / cytology
  • Hepatocytes / physiology*
  • Humans
  • Organ Specificity
  • Oxygen Consumption
  • Pharmaceutical Preparations / metabolism
  • Substrate Specificity
  • Time Factors


  • Pharmaceutical Preparations
  • Cytochrome P-450 Enzyme System
  • Aspartate Aminotransferases