The hiPSC line, Sigma 0028, was purchased from Sigma–Aldrich and maintained on coated plates with Corning Matrigel hESC‐Qualified Matrix (Corning, 734–1440) in Essential 8 Flex Medium (E8 Flex; Gibco, A2858501). The cells were passaged at 65% confluency using 0.1% Ethylenediaminetetraacetic acid (EDTA, Life Technologies, 15575020) in PBS. The hiPSCs were genetically engineered to induce the overexpression of 3 transcription factors (TFs), namely HNF1A, FOXA3, and PROX1 (named HC3X‐PSC) as described earlier to generate hepatocyte‐like progeny; or one TF, namely ETV2 (named iETV2‐PSC) to generate endothelial cells.[33 (link),
69 (link)
] When the cells reached 65% confluency, cells were dissociated into single cells using StemPro Accutase (Gibco, A2644501) cell dissociation reagent. Cells were plated on Matrigel Based Membrane Growth Factor Reduced coated plates (Corning, 734–0270) at a density of ±8.75 × 104 cells cm−2 or 3 × 105 cells mL−1 in mTeSR medium (Stem Cell Technologies, 85850) supplemented with 1:100 RevitaCell Supplement (Gibco, A2644501). When a cellular confluency of 70–80% was obtained, cells were differentiated toward either the hepatocyte or endothelial lineage. The use of hiPSCs for research was approved by the “Commissie Medische Ethiek,” UZ KU Leuven/Onderzoek U.Z. Gasthuisberg, Herestraat 49, B 3000 Leuven, under file no. S52426.
69 (link)
] When the cells reached 65% confluency, cells were dissociated into single cells using StemPro Accutase (Gibco, A2644501) cell dissociation reagent. Cells were plated on Matrigel Based Membrane Growth Factor Reduced coated plates (Corning, 734–0270) at a density of ±8.75 × 104 cells cm−2 or 3 × 105 cells mL−1 in mTeSR medium (Stem Cell Technologies, 85850) supplemented with 1:100 RevitaCell Supplement (Gibco, A2644501). When a cellular confluency of 70–80% was obtained, cells were differentiated toward either the hepatocyte or endothelial lineage. The use of hiPSCs for research was approved by the “Commissie Medische Ethiek,” UZ KU Leuven/Onderzoek U.Z. Gasthuisberg, Herestraat 49, B 3000 Leuven, under file no. S52426.
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