Day (0; or day 10 from iPSC): Isolated CD43+ iHPCs were washed using iMGL base differentiation medium and centrifuged at 300 × g for 6 minutes at room temperature. After centrifugation, supernatant was aspirated to waste and iHPCs were gently suspended in complete differentiation medium: M-CSF (25 ng/ml), IL-34 (100 ng/ml; Peprotech), and TGFβ-1 (50 ng/ml; Militenyi) added fresh each time. Cell density was adjusted to seed at density of 1–2 × 105 cells in 2 ml of complete medium per well in growth factor-reduced Matrigel-coated 6-well plates.
