Figure 1.

Hypertonic sodium-rich media decrease viability of human neutrophils and induce primary or secondary cell necrosis in vitro. Cell viability was assessed by flow cytometry after four-color death staining with PI, Hoechst33342, Annexin A5-FITC and DilC1(5) in various salt conditions (1) isotonicity 137 mM NaCl, (2) moderate hypertonicity 200 mM NaCl, and (3) high hypertonicity 300 mM NaCl, at various time points. (A) Flowcytometric dot plot analysis showing the gating strategy used to identify viable, apoptotic, and necrotic cell populations; (B) culture without (w/o) serum; (C) culture with (w/) 10% heat-inactivated FCS. Data were obtained from 3-5 healthy individuals and are presented as mean ± SD. Statistical analysis was performed using Mixed Effects Analysis. Table 1 displays all numerical P values. pNecrotic, primary necrotic; sNecrotic, secondary necrotic.