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

Figure 3

Effects of acute hypoxia on the development of giant phagocytes (Gϕ). Freshly isolated PMN were exposed for 24 h to intermittent hypoxia (IH, 56 cycles), sustained hypoxia (SH), or normoxia (N) and then cultured at normoxia for additional six days. Cytospins were prepared and analyzed by confocal microscopy (see Materials and Methods). Nuclei were stained with DAPI (blue). (a) Fixed cytospins were stained with anti-CD66b Abs and (b) neutrophil elastase (NE). (c) Isotype controls: fixed cytospins were stained with mouse IgG2 followed by 1/400 CF 647 goat anti-mouse IgG (red) staining. (d) Viable cells were stained with LysoTracker before fixation (see Materials and Methods). Arrow indicates large phagolysosomes. Representative data out of 3 independent experiments.

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