Fig. 1

Differentiation of iPS into Mφ. The upper panel is a schematic representation of the three-step EB-based protocol employed for hematopoietic differentiation. a Embryoid bodies (EB) are formed for 11 days; (b) EB started to gradually adhere in the presence of 25 ng/ml IL-3 with 50 ng/ml M-CSF added from day 12 onward; (c) Most EB were propagated in the presence of 25 ng/ml IL-3 with the addition of 50 ng/ml M-CSF from day 15 onward; (d) all EB were propagated and adherent in the presence of 25 ng/ml IL-3 with the addition of 50 ng/ml M-CSF from day 19 onward; (e-f) The cells were cultured for 25–30 days and were differentiated into monocytes (some round suspension cells were present), and suspension cells were collected every 4–5 days and changed to fresh medium; (g) collected suspension cells were cultured in the presence of 50 ng/ml IL-3 with 100 ng/ml M-CSF; (h) terminal differentiation is achieved by 35–40 days of adherent culture in the presence of 50 ng/ml IL-3 with 100 ng/ml M-CSF. The cells have typical macrophage morphological characteristics (round, oval, spindle and irregular, etc., and some cells showed pseudopods and convex forms)