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  • 1.
    Asplund, Annika
    et al.
    Högskolan i Skövde, Institutionen för biovetenskap. Högskolan i Skövde, Forskningscentrum för Systembiologi. Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden.
    Pradip, Arvind
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden / Novo Nordisk A/S, Bagsværd, Denmark.
    van Giezen, Mariska
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden.
    Aspegren, Anders
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden.
    Choukair, Helena
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden.
    Rehnström, Marie
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden.
    Jacobsson, Susanna
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden.
    Ghosheh, Nidal
    Högskolan i Skövde, Institutionen för biovetenskap. Högskolan i Skövde, Forskningscentrum för Systembiologi.
    El Hajjam, Dorra
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden.
    Holmgren, Sandra
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden / Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
    Larsson, Susanna
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden.
    Benecke, Jörg
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden.
    Butron, Mariela
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden.
    Wigander, Annelie
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden.
    Noaksson, Karin
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden.
    Sartipy, Peter
    Högskolan i Skövde, Institutionen för biovetenskap. Högskolan i Skövde, Forskningscentrum för Systembiologi. AstraZeneca R&D, GMD CVMD GMed, Mölndal, Sweden.
    Björquist, Petter
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden / NovaHep AB, Gothenburg, Sweden.
    Edsbagge, Josefina
    Takara Bio Europe AB (former Cellartis AB), Gothenburg, Sweden.
    Küppers-Munther, Barbara
    Högskolan i Skövde, Institutionen för biovetenskap. Högskolan i Skövde, Forskningscentrum för Systembiologi. Takara Bio Europe AB (former Cellartis AB), Arvid Wallgrens Backe 20, 413 46, Gothenburg, Sweden.
    One Standardized Differentiation Procedure Robustly Generates Homogenous Hepatocyte Cultures Displaying Metabolic Diversity from a Large Panel of Human Pluripotent Stem Cells2016Ingår i: Stem Cell Reviews, ISSN 1550-8943, E-ISSN 1558-6804, Vol. 12, nr 1, s. 90-104Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Human hepatocytes display substantial functional inter-individual variation regarding drug metabolizing functions. In order to investigate if this diversity is mirrored in hepatocytes derived from different human pluripotent stem cell (hPSC) lines, we evaluated 25 hPSC lines originating from 24 different donors for hepatic differentiation and functionality. Homogenous hepatocyte cultures could be derived from all hPSC lines using onestandardized differentiation procedure. To the best of our knowledge this is the first report of a standardized hepatic differentiation procedure that is generally applicable across a large panel of hPSC lines without any adaptations to individual lines. Importantly, with regard to functional aspects, such as Cytochrome P450 activities, we observed that hepatocytes derived from different hPSC lines displayed inter-individual variation characteristic for primary hepatocytes obtained from different donors, while these activities were highly reproducible between repeated experiments using the same line. Taken together, these data demonstrate the emerging possibility to compile panels of hPSC-derived hepatocytes of particular phenotypes/genotypes relevant for drug metabolism and toxicity studies. Moreover, these findings are of significance for applications within the regenerative medicine field, since our stringent differentiation procedure allows the derivation of homogenous hepatocyte cultures from multiple donors which is a prerequisite for the realization of future personalized stem cell based therapies.

  • 2.
    Asplund, Annika
    et al.
    Takara Bio Europe AB, Gothenburg, Sweden.
    Synnergren, Jane
    Högskolan i Skövde, Institutionen för biovetenskap. Högskolan i Skövde, Forskningscentrum för Systembiologi.
    Andersson, Christian X.
    Takara Bio Europe AB, Gothenburg, Sweden.
    Küppers-Munther, Barbara
    Takara Bio Europe AB, Gothenburg, Sweden.
    A novel maintenance medium extends the life-span and enables long term applications for both human primary hepatocytes and human pluripotent stem cell derived hepatocytes in conventional 2D cultures2017Konferensbidrag (Refereegranskat)
  • 3.
    Kia, Richard
    et al.
    Univ Liverpool, Dept Mol & Clin Pharmacol, MRC Ctr Drug Safety Sci, Liverpool, England.
    Kelly, Lorna
    Univ Liverpool, Dept Mol & Clin Pharmacol, England / Stem Cells Safer Med, London, England.
    Sison-Young, Rowena L. C.
    Univ Liverpool, Dept Mol & Clin Pharmacol, MRC Ctr Drug Safety Sci, Liverpool, England.
    Zhang, Fang
    Univ Liverpool, Dept Mol & Clin Pharmacol, England / Stem Cells Safer Med, London, England.
    Pridgeon, Chris S.
    Univ Liverpool, Dept Mol & Clin Pharmacol, England / Stem Cells Safer Med, London, England.
    Heslop, James A.
    Univ Liverpool, Dept Mol & Clin Pharmacol, MRC Ctr Drug Safety Sci, Liverpool, England.
    Metcalfe, Pete
    Univ Liverpool, Dept Mol & Clin Pharmacol, MRC Ctr Drug Safety Sci, Liverpool, England.
    Kitteringham, Neil R.
    Univ Liverpool, Dept Mol & Clin Pharmacol, England / Stem Cells Safer Med, London, England.
    Baxter, Melissa
    Univ Manchester, Fac Life Sci, Manchester, England / Univ Cent Lancashire, Sch Med & Dent, Preston, England.
    Harrison, Sean
    Stem Cells Safer Med, London, England / Acad Hlth Sci Ctr, Fac Med & Human Sci, Ctr Endocrinol & Diabet,Inst Human Dev, Manchester, England.
    Hanley, Neil A.
    Stem Cells Safer Med, London, England / Univ Manchester, Manchester Acad Hlth Sci Ctr, Fac Med & Human Sci, Ctr Endocrinol & Diabet,Inst Human Dev, Manchester, England / Cent Manchester Univ Hosp NHS Fdn Trust, Endocrinol Dept, Manchester England.
    Burke, Zoe D.
    Stem Cells Safer Med, London, England / Univ Bath, Dept Biol & Biochem, Ctr Regenerat Med, Bath, England.
    Storm,, Mike P.
    Stem Cells Safer Med, London, England / Univ Bath, Dept Biol & Biochem, Ctr Regenerat Med, Bath, England.
    Welham, Melanie J.
    Univ Bath, Dept Biol & Biochem, Ctr Regenerat Med, Bath, England.
    Tosh, David
    Stem Cells Safer Med, London, England / Univ Bath, Dept Biol & Biochem, Ctr Regenerat Med, Bath, England.
    Küppers-Munther, Barbara
    Högskolan i Skövde, Institutionen för biovetenskap. Högskolan i Skövde, Forskningscentrum för Systembiologi. Takara Bio Europe AB, Gothenburg, Sweden.
    Edsbagge, Josefina
    Takara Bio Europe AB, Gothenburg, Sweden.
    Lewis, Philip J. Starkey
    Univ Edinburgh, MRC Ctr Regenerat Med, Edinburgh EH16 4UU, Midlothian, Scotland.
    Bonner, Frank
    Stem Cells Safer Med, London, England.
    Harpur, Ernie
    Stem Cells Safer Med, London, England / Newcastle Univ, Inst Cellular Med, Sch Med, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England.
    Sidaway, James
    Univ Edinburgh, MRC Ctr Regenerat Med, Edinburgh EH16 4UU, Midlothian, Scotland / AstraZeneca R&D, Drug Safety & Metab, Cheshire, England.
    Bowes, Joanne
    Univ Edinburgh, MRC Ctr Regenerat Med, Edinburgh EH16 4UU, Midlothian, Scotland / AstraZeneca R&D, Drug Safety & Metab, Cheshire, England.
    Fenwick, Stephen W.
    Aintree Univ Hosp NHS Fdn Trust, North Western Hepatobiliary Unit, Liverpool, England.
    Malik, Hassan
    Aintree Univ Hosp NHS Fdn Trust, North Western Hepatobiliary Unit, Liverpool, England.
    Goldring, Chris E. P.
    Univ Liverpool, Dept Mol & Clin Pharmacol, MRC Ctr Drug Safety Sci, Liverpool, England / Stem Cells Safer Med, London England.
    Park, B. Kevin
    Univ Liverpool, Dept Mol & Clin Pharmacol, MRC Ctr Drug Safety Sci, Liverpool, England / Stem Cells Safer Med, London, England.
    MicroRNA-122: a novel hepatocyte-enriched in vitro marker of drug-induced cellular toxicity2015Ingår i: Toxicological Sciences, ISSN 1096-6080, E-ISSN 1096-0929, Vol. 144, nr 1, s. 173-185Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Emerging hepatic models for the study of drug-induced toxicity include pluripotent stem cell-derived hepatocyte-like cells (HLCs) and complex hepatocyte-non-parenchymal cellular coculture to mimic the complex multicellular interactions that recapitulate the niche environment in the human liver. However, a specific marker of hepatocyte perturbation, required to discriminate hepatocyte damage from non-specific cellular toxicity contributed by non-hepatocyte cell types or immature differentiated cells is currently lacking, as the cytotoxicity assays routinely used in in vitro toxicology research depend on intracellular molecules which are ubiquitously present in all eukaryotic cell types. In this study, we demonstrate that microRNA-122 (miR-122) detection in cell culture media can be used as a hepatocyte-enriched in vitro marker of drug-induced toxicity in homogeneous cultures of hepatic cells, and a cell-specific marker of toxicity of hepatic cells in heterogeneous cultures such as HLCs generated from various differentiation protocols and pluripotent stem cell lines, where conventional cytotoxicity assays using generic cellular markers may not be appropriate. We show that the sensitivity of the miR-122 cytotoxicity assay is similar to conventional assays that measure lactate dehydrogenase activity and intracellular adenosine triphosphate when applied in hepatic models with high levels of intracellular miR-122, and can be multiplexed with other assays. MiR-122 as a biomarker also has the potential to bridge results in in vitro experiments to in vivo animal models and human samples using the same assay, and to link findings from clinical studies in determining the relevance of in vitro models being developed for the study of drug-induced liver injury.

  • 4.
    Küppers-Munther, Barbara
    et al.
    Takara Bio Europe AB, Gothenburg, Sweden.
    Asplund, A.
    Takara Bio Europe AB, Gothenburg, Sweden.
    Ulfenborg, Benjamin
    Högskolan i Skövde, Institutionen för biovetenskap. Högskolan i Skövde, Forskningscentrum för Systembiologi.
    Synnergren, Jane
    Högskolan i Skövde, Institutionen för biovetenskap. Högskolan i Skövde, Forskningscentrum för Systembiologi.
    Abadie, A.
    Takara Bio Europe SAS, Paris, France.
    Novel human iPSC-derived hepatocytes with advanced functionality and long-term 2D cultures of human primary hepatocytes for metabolic disease studies2018Ingår i: Human Gene Therapy, ISSN 1043-0342, E-ISSN 1557-7422, Vol. 29, nr 12, s. A146-A146, artikel-id P406Artikel i tidskrift (Refereegranskat)
  • 5.
    Prokop, Andreas
    et al.
    Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester, United Kingdom.
    Küppers-Munther, Barbara
    Högskolan i Skövde, Institutionen för vård och natur. Högskolan i Skövde, Forskningscentrum för Systembiologi. Cellartis, Göteborg, Sweden.
    Sánchez-Soriano, Natalia
    Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester, United Kingdom.
    Using primary neuron cultures of Drosophila to analyze neuronal circuit formation and function2012Ingår i: The making and un-making of neuronal circuits in Drosophila / [ed] Bassem A. Hassan, Springer Science+Business Media B.V., 2012, s. 225-247Kapitel i bok, del av antologi (Refereegranskat)
    Abstract [en]

    For many decades, primary neuron cultures of Drosophila have been used complementary to work in vivo. Primary cultures were instrumental for the analysis of physiological properties of Drosophila neurons and synapses, and they were used for the analysis of developmental processes. Recent developments have established Drosophila primary neurons based on Schneider's culture media, as a means to investigate the neuronal cytoskeleton, opening up novel opportunities for research into cellular mechanisms of axonal growth, synapse formation and perhaps even neuronal degeneration. These cell cultures provide readouts for cytoskeletal dynamics that are difficult or impossible to access in vivo, and which turned out to be highly conserved with mammalian or other vertebrate neurons. Therefore, the same genetic manipulations in Drosophila can now be studied synergetically in culture and in vivo, to address cell biological principles of neuronal circuit formation and function. Here we describe in detail how these cell cultures are generated and discuss principal considerations for the experimental design and the solution of common problems. Furthermore, we describe in detail how to generate Schneider's media with adjustable inorganic ion concentrations. These media have been shown to promote the physiological maturation of neurons, thus expanding the use of the primary neuron cultures into the synaptic stage. The culture strategies described here recapitulate in vivo development with impressive accuracy and provide a promising means for Drosophila research on neuronal development and function.

  • 6.
    Ulvestad, Maria
    et al.
    AstraZeneca, Sweden / Cellectis AB, Sweden / University of Oslo, Norway.
    Nordell, Pär
    AstraZeneca, Sweden.
    Asplund, Annika
    Cellectis AB, Sweden.
    Rehnström, Marie
    Cellectis AB, Sweden.
    Jacobsson, Susanna
    Cellectis AB, Sweden.
    Holmgren, Gustav
    Högskolan i Skövde, Institutionen för vård och natur. Högskolan i Skövde, Forskningscentrum för Systembiologi. Cellectis AB, Sweden / Sahlgrenska University Hospital, Sweden.
    Davidson, Lindsay
    University of Dundee, Scotland.
    Brolén, Gabriella
    AstraZeneca, Sweden.
    Edsbagge, Josefina
    Cellectis AB, Sweden.
    Björquist, Petter
    Cellectis AB, Sweden.
    Küppers-Munther, Barbara
    Högskolan i Skövde, Institutionen för vård och natur. Högskolan i Skövde, Forskningscentrum för Systembiologi. Cellectis AB, Sweden.
    Andersson, Tommy B.
    AstraZeneca, Sweden / Karolinska Institute, Sweden.
    Drug metabolizing enzyme and transporter protein profiles of hepatocytes derived from human embryonic and induced pluripotent stem cells2013Ingår i: Biochemical Pharmacology, ISSN 0006-2952, E-ISSN 1356-1839, Vol. 86, nr 5, s. 691-702Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    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. (C) 2013 Elsevier Inc. All rights reserved.

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