Biochemical Engineering Institute, Saarland University, Saarbruecken, Germany.
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.
随着主要研究重点转向三维（3D）培养技术，中空纤维生物反应器由于允许在体外进行受控培养同时支持体内环境，因此显示出巨大的潜力，能够形成类似组织的结构。在这项研究中，我们在连续运行超过 2 周的 3D 中空纤维生物反应器系统中对人肝细胞进行了系统和详细的生理学表征。原代人肝细胞在整个培养过程中保持存活和功能。一般细胞功能，如氧摄取、氨基酸代谢和底物消耗，以及肝脏特异性功能，如药物代谢能力和肝脏特异性代谢产物的产生，均稳定超过 2 周。如预期的那样，在肝脏特异性功能方面观察到供体间的可变性，即尿素和白蛋白的产生。此外，我们在该设置中展示了在无血清条件下维持原代人肝细胞。细胞分离 3 周后稳定的基础细胞色素 P450 活性证明了这种系统在药理学应用中的潜力。该 3D 生物反应器系统中的肝细胞最终不仅可用于长期代谢和毒性研究，还可用于慢性重复剂量毒性评估。
