Anti cd45 coated beads

HSCs were generated from hiPSCs using STEMdiff^™ Hematopoietic Kit (StemCell Technologies), according to the manufacturer’s instructions. All cell culture in this method was performed under normoxic conditions. Cells harvested on day 12 were differentiated into iMACs based on either of following protocols: HSCs underwent magnetic sorting (MACS) using anti-CD34-coated beads (Miltenyi Biotec) and cultured for 7 days in RPMI (Corning) supplemented with 10% FBS, 1% penicillin/streptomycin (Thermo Fisher Scientific), and 100 ng/ml M-CSF (Peprotech). Harvested cells underwent further differentiation in the same media described above and were followed by MACS sorting using anti-CD45-coated beads (Miltenyi Biotec) on day 19. As the quality of HSCs harvested on day 12 was the most critical for generating well matured iMACs on day 19, we defined a differentiation as successful when both the harvested cell number on day 12 was more than 500,000 per well in a 12-well plate, and the cell viability was more than 70%. Once matured, 2D-iMACs were maintained under the same condition as described in 3D culture method or polarized into M1-like phenotype by culturing for 24 hours with the combination of 20 ng/ml IFN-γ (Peprotech) and 10 ng/ml LPS (MilliporeSigma).