Optimizing the Method for Differentiation of Macrophages from Human Induced Pluripotent Stem Cells

Figure 3

The hiPSC-derived macrophages possess the function of phagocytosis. (a, b) Flow cytometric analysis of the uptake of fluorescent beads by IPSDMs in two differentiation protocols. (c) Phagocytosis of fluorescent beads (red) by IPSDMs and IPSCs control. DAPI (blue) shows the cell nuclei; WGA (green) shows the cell membrane. Scale bars, 50 μm. (d) When the ratio of IPSDMs to tumor cells is 1 : 1, 1 : 2, or 1 : 4, the proportion of IPSDMs that have engulfed tumor cells is calculated. All experiments were performed three times. Error bars, SD. (e) Representative images of Nalm6 and Reh cells (labeled with Hoechst 33342) phagocytized by IPSDMs (labeled with CD11b and phase contrast image). Scale bars, 25 μm. (f) Flow cytometric analysis of phagocytosis of IPSDMs on Reh (). CD11b⁺ macrophages are gated (middle panel), and V450 intensity is shown as a histogram (right panel). Macrophages cocultured with Reh-Hoechst 33342 suspension (after washing) as a negative control.

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