PT  - JOURNAL ARTICLE
AU  - Hoekstra, Ruurdtje
AU  - Nibourg, Geert A.A.
AU  - van der Hoeven, Tessa V.
AU  - Plomer, Gabrielle
AU  - Seppen, Jurgen
AU  - Ackermans, Mariëtte T.
AU  - Camus, Sandrine
AU  - Kulik, Wim
AU  - van Gulik, Thomas M.
AU  - Elferink, Ronald P. Oude
AU  - Chamuleau, Robert A.F.M.
TI  - Phase 1 and Phase 2 Drug Metabolism and Bile Acid Production of HepaRG Cells in a Bioartificial Liver in Absence of Dimethyl Sulfoxide
AID  - 10.1124/dmd.112.049098
DP  - 2013 Mar 01
TA  - Drug Metabolism and Disposition
PG  - 562--567
VI  - 41
IP  - 3
4099  - http://dmd.aspetjournals.org/content/41/3/562.short
4100  - http://dmd.aspetjournals.org/content/41/3/562.full
SO  - Drug Metab Dispos2013 Mar 01; 41
AB  - The human liver cell line HepaRG has been recognized as a promising source for in vitro testing of metabolism and toxicity of compounds. However, currently the hepatic differentiation of these cells relies on exposure to dimethylsulfoxide (DMSO), which, as a side effect, has a cytotoxic effect and represses an all-round hepatic functionality. The AMC-bioartificial liver (AMC-BAL) is a three-dimensional bioreactor that has previously been shown to upregulate various liver functions of cultured cells. We therefore cultured HepaRG cells in the AMC-BAL without DMSO and characterized the drug metabolism. Within 14 days of culture, the HepaRG-AMC-BALs contained highly polarized viable liver-like tissue with heterogeneous expression of CYP3A4. We found a substantial metabolism of the tested substrates, ranging from 26% (UDP-glucuronosyltransferase 1A1), 47% (CYP3A4), to 240% (CYP2C9) of primary human hepatocytes. The CYP3A4 activity could be induced 2-fold by rifampicin, whereas CYP2C9 activity remained equally high. The HepaRG-AMC-BAL secreted bile acids at 43% the rate of primary human hepatocytes and demonstrated hydroxylation, conjugation, and transport of bile salts. Concluding, culturing HepaRG cells in the AMC-BAL yields substantial phase 1 and phase 2 drug metabolism, while maintaining high viability, rendering DMSO addition superfluous for the promotion of drug metabolism. Therefore, AMC-BAL culturing makes the HepaRG cells more suitable for testing metabolism and toxicity of drugs.