Matrigel coated 6 well plates

Cells were grown at 37 °C in a humidified environment under hypoxic conditions (5% O₂, 5% CO₂, and 90% N₂). Cells were cultured in mTeSR^TM1 (Stemcell Technologies, Cologne, Germany) on Matrigel^TM-coated 6-well plates (VWR, Darmstadt, Germany; 0.083 mg/well) in 1 mL Knockout^TM DMEM (Thermo Fisher Scientific, Bremen, Germany). For passaging, cells were treated with 1 mg/mL collagenase IV (Thermo Fisher Scientific) in Knockout DMEM for 5–7 min at 37 °C, followed by rinsing once in Knockout DMEM, and subsequently mechanical dissociation and seeding at a 1:10–1:100 split ratio with daily medium replacement.

HiPSCs were cultured at 37 °C, 95% humidity, and 5% CO₂. Cells were cultured in mTeSR^TM1 (Stemcell Technologies) supplemented with 1% gentamycin (Thermo Fisher Scientific) on Matrigel^TM-coated 6-well plates (VWR; 0.083 mg/well) in 1 ml Knockout™ DMEM (Thermo Fisher Scientific). For passaging, cells were treated with 1 mg/ml collagenase IV (Thermo Fisher Scientific) in Knockout DMEM for 30 min at 37 °C or Gentle Cell Dissociation Reagent (Stemcell Technologies) for 7 min at room temperature, followed by rinsing once in Knockout DMEM, and subsequently mechanical dissociation and seeding at a 1:10 − 1:100 split ratio with daily medium replacement.