Figure 5.

Influence of USSC on maturation of monocyte‐derived dendritic cells. CD14⁺ monocytes (2×10⁶) were cultured in the absence (Control, F) or presence of irradiated USSC at the cell numbers indicated (A) or 30,000 cells (B, C, D, E) for 6 days with GM‐CSF and IL‐4, followed by a 3‐day maturation period in the presence of GM‐CSF, IL‐4 and TNFα. After a total of 9 days of co‐culture, the frequency of CD83⁺ cells (A, B, C), the normalized relative expression (mean fluorescence of control cells was set to 1) of the molecules indicated (D) and forward (FSC)/side scatter (SSC) characteristics of cells (E, F) were determined by flow cytometry. Morphology was evaluated by differential interference contrast microscopy (E, F). Horizontal lines (B, D) indicate mean values. In certain experiments (C), USSC and DC in co‐cultures were separated by a membrane (Transwell), USSC conditioned medium (30%) was used instead of cells (USSC CM) or the activity of TGFβ was neutralized with a monoclonal antibody (+α‐TGF‐β; 5 μg/ml α‐TGF‐β(1, 2, 3)). Statistical significance is indicated (*–P≤ 0.05; **–P≤ 0.01).