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. 2020 May 28;21(11):3831. doi: 10.3390/ijms21113831

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Characterisation of induced pluripotent stem cell (iPSC)-derived hepatocyte-like cells (HLCs) on d20 of differentiation. (a) Fixed cells (P106-HLCs) were stained for albumin, AFP, and HNF4α to assess hepatic characteristics and maturation state of HLCs. Immunofluorescence staining (40x magnification) shows that d20 HLCs co-express albumin and AFP representing mature and fetal characteristics (five fields of view per well with 400 cells, three wells per condition). (b) Metabolic activity. The metabolic activity of P106- and 33D6-HLCs was determined by measuring the activity of cytochrome P450 (CYP)1A2 and CYP3A in HLCs. CYP-activity was measured in RLU and normalised to ml and mg of total protein (N = 4). d20 HLCs are metabolically active, showing successful differentiation. P106-HLCs show improved metabolic activity compared with 33D6-HLCs. Results are shown as mean +/− SEM. AFP = alpha fetoprotein, DAPI = 4′,6-Diamidin-2-phenylindol, HNF4α = hepatocyte nuclear factor 4 alpha, RLU = relative light units.