Differentiation of hESCs and hiPSCs into Germ Layers

H9 hESCs (WiCell, Madison, WI, USA) and the hiPSC lines FSPS13B, CF03, CF04, and CF05 were plated on vitronectin-coated plates (10 μg/mL, Stem Cell Technologies) and cultured in E6 media supplemented with 2 ng/mL transforming growth factor β (R&D) and 25 ng/mL fibroblast growth factor 2 (Dr. Marko Hyvönen, Cambridge University) making complete E8 media. Cells were maintained by weekly passaging using 0.5 mM EDTA (Thermo Fisher Scientific). The cells were differentiated into the three germ layers and functional cell types as previously described (Cheung et al., 2012 (link), Mendjan et al., 2014 (link)) and as described in the Supplemental Information.