Fig 1.

Differentiation of human monocytes into morphologically, phenotypically, and functionally distinct Mϕ subsets. (A) Monocytes were purified from buffy coats with a negative immunomagnetic selection and then incubated for 7 days in the presence of 100 ng/ml of either GM-CSF or M-CSF. The representative scanning electron microscopy pictures show the morphological features of monocytes immediately after isolation and monocyte-derived Mϕ at day 7 of in vitro culture. Bars, 10 μm. (B) Monocytes or M1 or M2 Mϕ were harvested, stained for the indicated markers and examined by flow cytometry. Bars depict mean values ± standard deviations (SD) for five blood donors. *, P < 0.05. (C) M1 or M2 Mϕ were seeded in fresh medium (1 × 10⁶ cells/ml) and either left untreated (n.s.) or stimulated for 24 h with LPS (100 ng/ml) and IFN-γ (20 ng/ml). The concentrations of the indicated cytokines/chemokines were evaluated by Bio-Plex technology. Each symbol represents cells obtained from one blood donor. Horizontal lines represent mean values ± standard errors of the means (SEM). *, P < 0.05.