Human neutrophil survival is extended while conditioned under anoxic conditions. (A) Neutrophil viability (Annexin V⁻/PI⁻) and absolute cell count were calculated on freshly purified neutrophils or after 20 hours incubation under anoxic conditions at 37°C in RPMI + HEPES (10 mM) with 3 mM glucose and 32 μg/mL DMOG supplementation. Error bars indicate standard deviation (SD). “ns” indicates P > .05 (n = 5). (B) Freshly purified and conditioned neutrophil morphology was assessed by transmission electron microscopy and compared with G-CSF–stimulated volunteer neutrophils (pseudo-Pelger-Huët phenotype). Bars represent 1 μm. The proportions of multilobed or bilobed neutrophils were calculated for each condition. (C) The cytosolic calcium flux was assessed on neutrophils described in panel B upon stimulation with fMLF (10 μM) in RPMI medium at 37°C, every 20 seconds. Error bars indicate SD. “ns” indicates P > .05 (n = 5).