Fig. 1
Generation and functional assay for macrophages from hPSCs. A Schematic overview of the stepwise direct hematopoietic and macrophage differentiation protocol from hPSCs. B Representative BF and Diff-Quick staining images of macrophages. Scale bars, 100 μm. C Phagocytosis of red fluorescein-labeled latex beads by hPSC-Macs. Scale bar, 100 μm. D Flow cytometry analysis of the expression of typical macrophage markers on hPSC-Macs. E The expression levels of inflammatory cytokines in macrophages after LPS stimulation for 24 h. Human macrophages obtained from BALF were used as a positive control. F Numbers of accumulated macrophages were counted every 3 days for up to 18 days, and the representative light microscopy images from different time points are shown. Scale bars, 100 μm. G CM and lysates were harvested from human PVCs and BM-MSC cultures and analyzed for expression of CyPA by western blot. Human skin fibroblasts (hFib) were used as a negative control. H Expression of CD147 in hPSC-derived hematopoietic progenitors was analyzed by flow cytometry. I Effects of CyPA on differentiation of macrophages. Hematopoietic progenitors were seeded at a density of 6 × 105 cells/well of 12 well plate and cultured in the presence and absence of CyPA (50 ng/mL) for 8 days. J Representative images of macrophages cultured in the presence and absence of CyPA. Scale bars, 20 μm. Data are presented as mean ± s.d. *p < 0.05, **p < 0.01