title: Drug metabolizing enzyme and transporter protein profiles of hepatocytes derived from human embryonic and induced pluripotent stem cells.
abstract: Human embryonic and induced pluripotent stem cell-derived hepatocytes (hESC-Hep and hiPSC-Hep) have the potential to provide relevant human in vitro model systems for toxicity testing and drug discovery studies. In this study, the expression and function of important drug metabolizing cytochrome P450 (CYP) enzymes and transporter proteins in hESC-Hep and hiPSC-Hep were compared to cryopreserved human primary hepatocytes (hphep) and HepG2 cells. Overall, CYP activities in hESC-Hep and hiPSC-Hep were much lower than in hphep cultured for 4 h, but CYP1A and 3A activities were comparable to levels in hphep cultured for 48h (CYP1A: 35% and 26% of 48 h hphep, respectively; CYP3A: 80% and 440% of 48 h hphep, respectively). Importantly, in hESC-Hep and hiPSC-Hep, CYP activities were stable or increasing for at least one week in culture which was in contrast to the rapid loss of CYP activities in cultured hphep between 4 and 48 h after plating. With regard to transporters, in hESC-Hep and hiPSC-Hep, pronounced NTCP activity (17% and 29% of 4 h hphep, respectively) and moderate BSEP activity (6% and 8% of 4 h hphep, respectively) were observed. Analyses of mRNA expression and immunocytochemistry supported the observed CYP and transporter activities and showed expression of additional CYPs and transporters. In conclusion, the stable expression and function of CYPs and transporters in hESC-Hep and hiPSC-Hep for at least one week opens up the possibility to reproducibly perform long term and extensive studies, e.g. chronic toxicity testing, in a stem cell-derived hepatic system.
authors.affiliation: DMPK, AstraZeneca R&D Mölndal, Pepparedsleden 1, SE-431 83 Mölndal, Sweden. maria.ulvestad@farmasi.uio.no
authors.forename: Maria
authors.initials: M
authors.lastname: Ulvestad
authors.forename: Pär
authors.initials: P
authors.lastname: Nordell
authors.forename: Annika
authors.initials: A
authors.lastname: Asplund
authors.forename: Marie
authors.initials: M
authors.lastname: Rehnström
authors.forename: Susanna
authors.initials: S
authors.lastname: Jacobsson
authors.forename: Gustav
authors.initials: G
authors.lastname: Holmgren
authors.forename: Lindsay
authors.initials: L
authors.lastname: Davidson
authors.forename: Gabriella
authors.initials: G
authors.lastname: Brolén
authors.forename: Josefina
authors.initials: J
authors.lastname: Edsbagge
authors.forename: Petter
authors.initials: P
authors.lastname: Björquist
authors.forename: Barbara
authors.initials: B
authors.lastname: Küppers-Munther
authors.forename: Tommy B
authors.initials: TB
authors.lastname: Andersson
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics / Q000378:metabolism; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000378:metabolism*; D022781:Hepatocytes / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics / Q000378:metabolism; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D027702:Organic Cation Transporter 1 / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D003577:Cytochrome P-450 Enzyme System; D026901:Membrane Transport Proteins; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D009363:Neoplasm Proteins; D027702:Organic Cation Transporter 1; C513055:ABCB1 protein, human; C000715087:ABCC2 protein, human; C116614:ABCG2 protein, human
keywords: BCRP; BSEP; BSP; CYP; Cytochrome P450; E17βG; E3S; Hepatocytes; Human embryonic stem cell-derived hepatocytes; Human induced pluripotent stem cell-derived hepatocytes; MDR1; MRP2; NTCP; OATP1B1; OCT1; RIT; TCA; Transporter proteins; bile salt export pump; breast cancer resistance protein; bromosulfophtalein; cytochrome P450; day 29 or 36; estradiol-17β-d-glucuronide; estrone-3-sulfate; hESC; hESC-Hep; hepatocyte cultures 29 or 36 days respectively after start of the hepatic differentiation protocol; hiPSC; hiPSC-Hep; hphep; human embryonic stem cell-derived hepatocytes; human embryonic stem cells; human induced pluripotent stem cell-derived hepatocytes; human induced pluripotent stem cells; human primary hepatocytes; multidrug resistance protein 1; multidrug resistance protein 2; organic anion-transporting polypeptide 1B1; organic cation transporter 1; ritonavir; sodium-taurocholate cotransporting polypeptide; taurocholic acid