FIG. 1.

Flow cytometric gating strategy and phenotypical characterization of monocyte-derived human dendritic cells. Purified human CD14+ monocytes were cultured for 5 days in the presence of IL-4 and granulocyte-macrophage colony-stimulating factor to generate immature dendritic cells (iDC). Subsequent lipopolysaccharide (LPS) stimulation of iDC for 24 h generated mature DC (iDC+LPS). Gating was performed as follows: (a) DCs were pregated according to forward and sideward scatter, (b) verified viable, (c) and CD14-negative before (d) classification as immature (CD14− CD86+ CD83−) or mature DC (CD14− CD86+ CD83+). (e) Histograms show the expression of surface markers CD86 and CD83 (black lines) in comparison to unlabeled cells (solid gray curves) for untreated (iDC) or iDC treated with LPS (iDC+LPS), chitosan (500 μg/mL; iDC+Ch500), agarose (500 μg/mL; iDC+Ag500), and for porcine collagen (500 μg/mL; iDC+pC500). Representative data from 5 independent experiments are shown.