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Analysis of drug metabolism activities in a miniaturized liver cell bioreactor for use in pharmacological studies

Abstract:

Based on a hollow fiber perfusion technology with internal oxygenation, a miniaturized bioreactor with a volume of 0.5 mL for in vitro studies was recently developed. Here, the suitability of this novel culture system for pharmacological studies was investigated, focusing on the model drug diclofenac. Primary human liver cells were cultivated in bioreactors and in conventional monolayer cultures in parallel over 10 days. From day 3 on, diclofenac was continuously applied at a therapeutic concentration (6.4 µM) for analysis of its metabolism. In addition, the activity and gene expression of the cytochrome P450 (CYP) isoforms CYP1A2, CYP2B6, CYP2C9, CYP2D6, and CYP3A4 were assessed. Diclofenac was metabolized in bioreactor cultures with an initial conversion rate of 230 ± 57 pmol/h/10(6) cells followed by a period of stable conversion of about 100 pmol/h/10(6) cells. All CYP activities tested were maintained until day 10 of bioreactor culture. The expression of corresponding mRNAs correlated well with the degree of preservation. Immunohistochemical characterization showed the formation of neo-tissue with expression of CYP2C9 and CYP3A4 and the drug transporters breast cancer resistance protein (BCRP) and multidrug resistance protein 2 (MRP2) in the bioreactor. In contrast, monolayer cultures showed a rapid decline of diclofenac conversion and cells had largely lost activity and mRNA expression of the assessed CYP isoforms at the end of the culture period. In conclusion, diclofenac metabolism, CYP activities and gene expression levels were considerably more stable in bioreactor cultures, making the novel bioreactor a useful tool for pharmacological or toxicological investigations requiring a highly physiological in vitro representation of the liver. Biotechnol. Bioeng. © 2012 Wiley Periodicals, Inc.

22688505

Projects: C2: Organization of the sinusoidal system and the liver lobule - referen...

Biotechnology and bioengineering
Biotechnol. Bioeng. 109(12): 3172-81
13th Jun 2012

Stefan A Hoffmann, Ursula Müller-Vieira, Klaus Biemel, Daniel Knobeloch, Sandra Heydel, Marc Lübberstedt, Andreas K Nüssler, Tommy B Andersson, Jörg C Gerlach, Katrin Zeilinger

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[Stefan Hoffmann] [Daniel Knobeloch] [Marc Lübberstedt] [Andreas Nüssler] [Katrin Zeilinger]

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Intercellular
Views: 2365
  • Created: 24th Aug 2012 at 11:07
  • Last updated: 24th Oct 2013 at 16:16

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