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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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 one standardized 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.
KeywordsHepatocyte differentiation Human induced pluripotent stem cells Human embyronic stem cells Liver Toxicity Cellular therapy
We thank other members of Takara Bio Europe AB and Skövde University who have provided assistance and technical help through-out this project.
Supported by the IMI-JU project MIP-DILI (grant 115336), the Seventh Framework Program project InnovaLiv (grant 278152), and the Knowledge Foundation (grants 2010/0069, 2012/0310, 2013/89).
Disclosure of Potential Conflicts of Interest
All authors are or have been employed by Takara Bio Europe AB (former Cellartis AB) when this study was performed.
Annika Asplund: Collection, assembly, analysis, and interpretation of data, manuscript writing.
Anders Aspegren: Conception and design, data analysis and interpretation.
Arvind Pradip, Mariska van Giezen, Marie Rehnström, Susanna Jacobsson, Nidal Ghosheh, Dorra El Hajjam, Sandra Holmgren, Susanna Larsson, Jörg Benecke, Mariela Butron, Annelie Wigander: Collection, assembly and analysis of data.
Helena Choukair, Karin Noaksson: Analysis and interpretation of data.
Peter Sartipy, Josefina Edsbagge: Interpretation of data, manuscript writing.
Petter Björquist: Conception and design, manuscript writing.
Barbara Küppers-Munther: Conception and design, assembly, analysis, and interpretation of data, manuscript writing.
All: Final approval of manuscript
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