Drug metabolizing enzyme and transporter protein profiles of hepatocytes derived from human embryonic and induced pluripotent stem cells

Maria Ulvestad, Pär Nordell, Annika Asplund, Marie Rehnström, Susanna Jacobsson, Gustav Holmgren, Lindsay Davidson, Gabriella Brolén, Josefina Edsbagge, Petter Björquist, Barbara Küppers-Munther, Tommy B. Andersson

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    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 48 h (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.
    Original languageEnglish
    Pages (from-to)691-702
    Number of pages12
    JournalBiochemical Pharmacology
    Volume86
    Issue number5
    DOIs
    Publication statusPublished - 1 Sep 2013

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    Induced Pluripotent Stem Cells
    Stem cells
    Hepatocytes
    Cytochrome P-450 Enzyme System
    Enzymes
    Pharmaceutical Preparations
    Proteins
    Toxicity
    Cytochrome P-450 CYP3A
    Hep G2 Cells
    Testing
    Drug Discovery
    Plating
    Human Embryonic Stem Cells
    Stem Cells
    Immunohistochemistry
    Messenger RNA

    Cite this

    Ulvestad, M., Nordell, P., Asplund, A., Rehnström, M., Jacobsson, S., Holmgren, G., ... Andersson, T. B. (2013). Drug metabolizing enzyme and transporter protein profiles of hepatocytes derived from human embryonic and induced pluripotent stem cells. Biochemical Pharmacology, 86(5), 691-702. https://doi.org/10.1016/j.bcp.2013.06.029
    Ulvestad, Maria ; Nordell, Pär ; Asplund, Annika ; Rehnström, Marie ; Jacobsson, Susanna ; Holmgren, Gustav ; Davidson, Lindsay ; Brolén, Gabriella ; Edsbagge, Josefina ; Björquist, Petter ; Küppers-Munther, Barbara ; Andersson, Tommy B. / Drug metabolizing enzyme and transporter protein profiles of hepatocytes derived from human embryonic and induced pluripotent stem cells. In: Biochemical Pharmacology. 2013 ; Vol. 86, No. 5. pp. 691-702.
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    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 48 h (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.",
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    Ulvestad, M, Nordell, P, Asplund, A, Rehnström, M, Jacobsson, S, Holmgren, G, Davidson, L, Brolén, G, Edsbagge, J, Björquist, P, Küppers-Munther, B & Andersson, TB 2013, 'Drug metabolizing enzyme and transporter protein profiles of hepatocytes derived from human embryonic and induced pluripotent stem cells', Biochemical Pharmacology, vol. 86, no. 5, pp. 691-702. https://doi.org/10.1016/j.bcp.2013.06.029

    Drug metabolizing enzyme and transporter protein profiles of hepatocytes derived from human embryonic and induced pluripotent stem cells. / Ulvestad, Maria; Nordell, Pär; Asplund, Annika; Rehnström, Marie; Jacobsson, Susanna; Holmgren, Gustav; Davidson, Lindsay; Brolén, Gabriella; Edsbagge, Josefina; Björquist, Petter; Küppers-Munther, Barbara; Andersson, Tommy B.

    In: Biochemical Pharmacology, Vol. 86, No. 5, 01.09.2013, p. 691-702.

    Research output: Contribution to journalArticle

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    T1 - Drug metabolizing enzyme and transporter protein profiles of hepatocytes derived from human embryonic and induced pluripotent stem cells

    AU - Ulvestad, Maria

    AU - Nordell, Pär

    AU - Asplund, Annika

    AU - Rehnström, Marie

    AU - Jacobsson, Susanna

    AU - Holmgren, Gustav

    AU - Davidson, Lindsay

    AU - Brolén, Gabriella

    AU - Edsbagge, Josefina

    AU - Björquist, Petter

    AU - Küppers-Munther, Barbara

    AU - Andersson, Tommy B.

    PY - 2013/9/1

    Y1 - 2013/9/1

    N2 - 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 48 h (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.

    AB - 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 48 h (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.

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