Effect of culture conditions on the differentiation of CD1a⁻ and CD1a⁺ monocyte-derived dendritic cells. Monocytes of a given individual were differentiated to DCs under different conditions. The results shown are representative of 4 independent experiments. (A-B) DCs were differentiated in various culture media as indicated: AIMV, or RPMI without or with 10% FCS, 10% LPDS, 10% AB serum, or 1% ITS. The percentage of CD1a⁺ cells was determined at day 5 of culture by flow cytometry. (C) DCs were cultured in RPMI medium substituted with graded concentration of LPDS (□), FCS (⊡), and AB serum (▪). RPMI without serum are represented as ▧. (D) DCs were differentiated in 10% LPDS (□) substituted with graded concentration of FCS (⊡) or AB serum (▪). (E) MoDCs were differentiated in RPMI+1% ITS in the presence of 40 μg/mL (▪) or 10 μg/mL (⊡) lipoproteins as indicated. Control cultures did not contain lipoproteins. (F) MoDCs were differentiated in different culture media as indicated, and the percentage of CD1a⁺ cells was measured in live (extracellular [ec] CD1a; □) and permeabilized cells (extra- and intracellular [ec + ic] CD1a; ⊡).