Elplasia six well plates

The hiPSCs (Ff-I01s04) were maintained on Laminin 511 E8 fragment-coated (iMatrix-511, kindly provided by Nippi, Incorporated, Tokyo, Japan) dishes in StemFit AK02N (Ajinomoto, Tokyo, Japan). The hiPSCs were differentiated into Hepatic endoderm (HE), endothelial (EC), and mesenchymal cells (MC), as described previously [25 (link)].
The generation of hiPSC-derived liver buds was carried out as described previously [13 (link)]. We collected HE, EC, and MC and seeded them on Elplasia six-well plates (Corning, Corning, NY, USA). The seeded cell number for HE was 2.5 × 10⁶ cells, for EC, 1 × 10⁶ cells, and, for MC, 1 × 10⁶ cells per well. The culture medium used was the same as reported previously [25 (link)]. Y-27632 (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) was added on day one. On day two, small liver buds were collected and reseeded on a cell culture insert (Corning, Corning, NY, USA). The medium was changed every other day.

The generation of human iPSC-derived liver organoids was carried out as described previously [20 ]. We collected hepatic endoderm, endothelial, and mesenchymal cells and seeded them on Elplasia six-well plates (Corning, Corning, NY, USA). The seeded cell number for hepatic endoderm was 2.5 × 10⁶ cells, for endothelial, 1 × 10⁶ cells, and, for mesenchymal cells, 1 × 10⁶ cells per well. The culture medium used was the same as reported previously [21 ]. Y-27632 (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) was added on day one. On day two, small liver organoids were collected and reseeded on a cell culture insert (Corning, Corning, NY, USA). The medium was changed every other day.