The hiPSC line Edi042A was purchased from Cedars Sinai. The hiPSC line WTC-ZO1-mEGFP (AICS-0023)27 (link) was a gift from the Gordana Vunjak-Novakovic lab. Both cell lines were maintained in six-well plates coated with Matrigel growth factor reduced basement membrane matrix (GFR-Matrigel, Corning, 354230) in mTeSR plus medium (STEMCELL Technologies, 100–0276). To coat the six-well plates, 1.5 mL of GFR-Matrigel (diluted at a 1:100 ratio with DMEM/F12 medium, Gibco, 11320033) was added per well and incubated at 37°C for 1h. To passage hiPSCs (60–70% confluency), cells were rinsed with 3–4 mL of DMEM/F12 medium per well, and then 2 mL of Accutase (Sigma-Aldrich, A6964) was added for 5–6 min at 37°C. Subsequently, 2 ml of mTeSR plus medium was added to neutralize the dissociation. Cells were collected after centrifugation at 1000 rpm for 3 min. Cells were resuspended with mTeSR plus with 10 μM Y27632 (Selleckchem, S1049) and evenly distributed on GFR-Matrigel coated wells.