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Küppers-Munther, BarbaraORCID iD iconorcid.org/0000-0003-4217-9355
Publications (5 of 5) Show all publications
Küppers-Munther, B., Asplund, A., Ulfenborg, B., Synnergren, J. & Abadie, A. (2018). Novel human iPSC-derived hepatocytes with advanced functionality and long-term 2D cultures of human primary hepatocytes for metabolic disease studies. Paper presented at Conference on Changing the Face of Modern Medicine - Stem Cell and Gene Therapy, OCT 16-19, 2018, Lausanne, SWITZERLAND. Human Gene Therapy, 29(12), A146-A146, Article ID P406.
Open this publication in new window or tab >>Novel human iPSC-derived hepatocytes with advanced functionality and long-term 2D cultures of human primary hepatocytes for metabolic disease studies
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2018 (English)In: Human Gene Therapy, ISSN 1043-0342, E-ISSN 1557-7422, Vol. 29, no 12, p. A146-A146, article id P406Article in journal, Meeting abstract (Refereed) Published
Place, publisher, year, edition, pages
USA: Mary Ann Liebert, 2018
National Category
Cell Biology
Research subject
Bioinformatics
Identifiers
urn:nbn:se:his:diva-16701 (URN)000453707700464 ()
Conference
Conference on Changing the Face of Modern Medicine - Stem Cell and Gene Therapy, OCT 16-19, 2018, Lausanne, SWITZERLAND
Available from: 2019-03-14 Created: 2019-03-14 Last updated: 2019-05-10Bibliographically approved
Asplund, A., Pradip, A., van Giezen, M., Aspegren, A., Choukair, H., Rehnström, M., . . . Küppers-Munther, B. (2016). One Standardized Differentiation Procedure Robustly Generates Homogenous Hepatocyte Cultures Displaying Metabolic Diversity from a Large Panel of Human Pluripotent Stem Cells. Stem Cell Reviews, 12(1), 90-104
Open this publication in new window or tab >>One Standardized Differentiation Procedure Robustly Generates Homogenous Hepatocyte Cultures Displaying Metabolic Diversity from a Large Panel of Human Pluripotent Stem Cells
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2016 (English)In: Stem Cell Reviews, ISSN 1550-8943, E-ISSN 1558-6804, Vol. 12, no 1, p. 90-104Article in journal (Refereed) Published
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.

Place, publisher, year, edition, pages
Springer, 2016
Keywords
Cellular therapy, Hepatocyte differentiation, Human embyronic stem cells, Human induced pluripotent stem cells, Liver, Toxicity
National Category
Cell and Molecular Biology
Research subject
Medical sciences; Bioinformatics
Identifiers
urn:nbn:se:his:diva-11781 (URN)10.1007/s12015-015-9621-9 (DOI)000374582000008 ()26385115 (PubMedID)2-s2.0-84955335024 (Scopus ID)
Available from: 2015-12-31 Created: 2015-12-31 Last updated: 2018-07-31Bibliographically approved
Kia, R., Kelly, L., Sison-Young, R. L. C., Zhang, F., Pridgeon, C. S., Heslop, J. A., . . . Park, B. K. (2015). MicroRNA-122: a novel hepatocyte-enriched in vitro marker of drug-induced cellular toxicity. Toxicological Sciences, 144(1), 173-185
Open this publication in new window or tab >>MicroRNA-122: a novel hepatocyte-enriched in vitro marker of drug-induced cellular toxicity
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2015 (English)In: Toxicological Sciences, ISSN 1096-6080, E-ISSN 1096-0929, Vol. 144, no 1, p. 173-185Article in journal (Refereed) Published
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.

Keywords
bridging biomarker, cell-specific biomarker, cytotoxicity, drug-induced liver injury, hepatocytes, in vitro model, microRNA
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:his:diva-13566 (URN)10.1093/toxsci/kfu269 (DOI)000353543000019 ()25527335 (PubMedID)2-s2.0-84924455476 (Scopus ID)
Available from: 2017-05-19 Created: 2017-05-19 Last updated: 2019-01-16Bibliographically approved
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
Open this publication in new window or tab >>Drug metabolizing enzyme and transporter protein profiles of hepatocytes derived from human embryonic and induced pluripotent stem cells
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2013 (English)In: Biochemical Pharmacology, ISSN 0006-2952, E-ISSN 1356-1839, Vol. 86, no 5, p. 691-702Article in journal (Refereed) Published
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.

Place, publisher, year, edition, pages
Elsevier, 2013
Keywords
Human embryonic stem cell-derived hepatocytes, Human induced pluripotent stem cell- derived hepatocytes, Cytochrome P450, Transporter proteins, Hepatocytes
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject
Natural sciences
Identifiers
urn:nbn:se:his:diva-8597 (URN)10.1016/j.bcp.2013.06.029 (DOI)000323404500012 ()23856292 (PubMedID)2-s2.0-84884903479 (Scopus ID)
Available from: 2013-10-31 Created: 2013-10-30 Last updated: 2017-12-06Bibliographically approved
Prokop, A., Küppers-Munther, B. & Sánchez-Soriano, N. (2012). Using primary neuron cultures of Drosophila to analyze neuronal circuit formation and function. In: Bassem A. Hassan (Ed.), The making and un-making of neuronal circuits in Drosophila: (pp. 225-247). Springer Science+Business Media B.V.
Open this publication in new window or tab >>Using primary neuron cultures of Drosophila to analyze neuronal circuit formation and function
2012 (English)In: The making and un-making of neuronal circuits in Drosophila / [ed] Bassem A. Hassan, Springer Science+Business Media B.V., 2012, p. 225-247Chapter in book (Refereed)
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.

Place, publisher, year, edition, pages
Springer Science+Business Media B.V., 2012
Series
Neuromethods, ISSN 0893-2336 ; 69
Research subject
Natural sciences
Identifiers
urn:nbn:se:his:diva-5818 (URN)10.1007/978-1-61779-830-6_10 (DOI)2-s2.0-84867547076 (Scopus ID)1-61779-829-0 (ISBN)978-1-61779-830-6 (ISBN)
Available from: 2012-05-02 Created: 2012-05-02 Last updated: 2017-11-27Bibliographically approved
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ORCID iD: ORCID iD iconorcid.org/0000-0003-4217-9355

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