Figure 2

Schematic representation of the iPSC differentiation protocol toward megakaryocytes (MKs) and platelets (PLTs). iPSCs maintained in StemMACS medium were seeded at a density of 50,000 cells/cm² on Laminin-521-coated plates 1 d before starting the differentiation. From d 0 to 2, cells were cultured in ½ StemMACS medium and ½ APEL medium containing vascular endothelial growth factor (VEGF, 50 ng/mL) and bone morphogenetic protein 4 (BMP4, 50 ng/mL). On d 2, the medium was changed to APEL medium containing VEGF and BMP4 (50 ng/mL). Cytokines were changed from d 4 on to thrombopoietin (TPO, 50 ng/mL), stem cell factor (SCF, 50 ng/mL) and interleukin-3 (IL-3, 25 ng/mL). On d 12, the medium was changed to APEL medium containing TPO and SCF (50 ng/mL). Half of the medium was replaced twice a week. Dotted arrows indicate complete media changes due to modification in cytokine composition.