Culturing human iPSCs on Laminin-511

The hiPSC lines (Tic and 1383D2) were obtained from the Japanese Collection of Research Bioresources Cell Bank and the Center for iPS Cell Research and Application at the Kyoto University [21 (link)], respectively. hiPSCs were cultured on polystyrene plates coated with laminin-511 E8 fragments (iMatrix-511; Nippi, Inc., Tokyo, Japan) in specific medium (StemFit AK02N medium; Ajinomoto, Tokyo, Japan). Single cells were seeded as 7.5 × 10³ cells/cm² with 10 μM ROCK inhibitor (CultureSure Y-27632; Fujifilm Wako Pure Chemical Corp., Osaka, Japan) at 37 °C in a humidified atmosphere with 5% CO₂. The culture medium was changed every 24 h.