Culturing hiPSC Line 1383D2 on Laminin-511

The hiPSC line, 1383D2, was obtained from the Center for iPS Cell Research and Application, Kyoto University (Kyoto, Japan). Cells were maintained on culture substrates coated with recombinant laminin-511 E8 fragments (iMatrix-511; Nippi, Inc., Tokyo, Japan) in a chemically defined and animal component-free medium (StemFit AK02N; Ajinomoto Co., Inc., Tokyo, Japan) following a previously published protocol [43 (link)]. The cells were cultured at a viable cell density of 7.5 × 10³ cells/cm² and subcultured every four days using TrypLE Select. For the first 24 h, 10 μM Rho-associated coiled-coil containing protein kinase (ROCK) inhibitor (Y-27632; Wako Pure Chemical Industries, Osaka, Japan) was used to enhance single hiPSC survival. The cells were incubated at 37 °C in a humidified atmosphere containing 5% CO₂, and the culture medium was replaced daily with fresh medium.