Figure 4.
Analysis of Migratory Potential of hiPSC-Derived Neutrophils
(A) Images from 45-min time-lapse movies of hiPSC-derived neutrophils during chemotaxis to vehicle control, fMLP, or IL-8 on fibrinogen. Yellow lines showing automated cell tracking. Scale bars, 100 μm.
(B) Quantification of chemotactic index and velocity of hiPSC neutrophils, showing significant differences of hiPSC neutrophils exposed to chemoattractant compared with treatment with vehicle control. p values for chemotactic index, control to IL-8 p = 0.0040, control to fMLP p < 0.0001. p values for velocity, control to IL-8 p = 0.0404, control to fMLP p = 0.0009 as determined by unpaired t test. Primary blood neutrophils showing chemotactic and velocity values. Bars show mean ± SE from 3 independent experiments with technical replicates, but not biological replicates for statistical analysis.
(C) Montage image from high-resolution time-lapse movie of hiPSC neutrophils migrating and phagocytosing Pseudomonas aeruginosa. Arrow indicates one point during the movie when the cell is targeting bacteria. Scale bars, 10 μm. All functional assays with hiPSC-derived neutrophils were performed on batches obtained from myeloid progenitors collected at 8 or 12 days after ETV2 transfection.