Intermittent Hypoxia Affects the Spontaneous Differentiation In Vitro of Human Neutrophils into Long-Lived Giant Phagocytes

Figure 1

Effects of cytochalasin B and GM-CSF/IL-4 on giant phagocyte (Gϕ) formation. (a) Freshly isolated PMN were incubated at time 0 without or with 10 μM cytochalasin B and followed for 7 days in culture. Bright-field microscopy of 7-day living cultures. (b) Representative photomicrographs of May-Grünwald-Giemsa-stained cytospin preparations of PMN cultured without (Gϕ) or with GM-CSF/IL-4 for 7 days. Samples were analyzed with a bright-field microscopy. Magnification, ×40. Cells developed in cultures with medium supplemented with GM-CSF/IL-4 show widespread cytoplasmic projections. Representative data out of 3 independent experiments. (c) PKH-26 (red) dye labeled PMN were cultured in cytokine-free medium for 7 days and PKH-67 (green) dye labeled PMN were cultured in medium supplemented with GM-CSF/IL-4 for 7 days. Then the developed cells were mixed in a ratio of 1 : 1 and cocultured for 2 h. Cytospins were fixed and analyzed by confocal microscopy.

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