title: Phase 1 and phase 2 drug metabolism and bile acid production of HepaRG cells in a bioartificial liver in absence of dimethyl sulfoxide.
abstract: 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.
authors.affiliation: Tytgat Institute for Liver and Intestinal Research, Amsterdam, The Netherlands. r.hoekstra@amc.uva.nl
authors.forename: Ruurdtje
authors.initials: R
authors.lastname: Hoekstra
authors.forename: Geert A A
authors.initials: GA
authors.lastname: Nibourg
authors.forename: Tessa V
authors.initials: TV
authors.lastname: van der Hoeven
authors.forename: Gabrielle
authors.initials: G
authors.lastname: Plomer
authors.forename: Jurgen
authors.initials: J
authors.lastname: Seppen
authors.forename: Mariëtte T
authors.initials: MT
authors.lastname: Ackermans
authors.forename: Sandrine
authors.initials: S
authors.lastname: Camus
authors.forename: Wim
authors.initials: W
authors.lastname: Kulik
authors.forename: Thomas M
authors.initials: TM
authors.lastname: van Gulik
authors.forename: Ronald P Oude
authors.initials: RP
authors.lastname: Elferink
authors.forename: Robert A F M
authors.initials: RA
authors.lastname: Chamuleau
mesh_terms: D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism; D001647:Bile Acids and Salts / Q000378:metabolism*; D001663:Bilirubin / Q000378:metabolism; D019149:Bioreactors; D001711:Biotransformation; D018929:Cell Culture Techniques; D002460:Cell Line; D065729:Cytochrome P-450 CYP2C9; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology* / Q000633:toxicity; D014453:Glucuronosyltransferase / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D006900:Hydroxylation; D007527:Isoenzymes; D007700:Kinetics; D019164:Liver, Artificial; D050216:Metabolic Detoxication, Phase I; D050217:Metabolic Detoxication, Phase II; D012293:Rifampin / Q000494:pharmacology; D013379:Substrate Specificity; D013739:Testosterone; D014044:Tolbutamide / Q000378:metabolism; D000099148:UGT1A1 Enzyme
chemical_list: D001189:Aryl Hydrocarbon Hydroxylases; D001647:Bile Acids and Salts; D001663:Bilirubin; D065729:Cytochrome P-450 CYP2C9; D051544:Cytochrome P-450 CYP3A; D004121:Dimethyl Sulfoxide; D014453:Glucuronosyltransferase; D007527:Isoenzymes; D012293:Rifampin; D013739:Testosterone; D014044:Tolbutamide; D000099148:UGT1A1 Enzyme; C450260:CYP2C9 protein, human; C510163:CYP3A4 protein, human