Generation of mature DCs by culturing murine Lin⁻c-kit⁺ HPCs in the presence of mouse TNFα, GM-CSF, and SCF. Morphologic analyses were performed on cultured cells (A through D) or Giemsa-stained cells after cytocentrifugation (E through H). (A through D) A phase contrast microscopic observation was performed on murine Lin⁻c-kit⁺ HPCs stimulated by the combination of GM-CSF + SCF (A) or that of GM-CSF + SCF + mouse TNFα (B through D) for 5 (A and B) or 14 days (C) as described in Fig 1. In (D), the aggregates shown in (C) were isolated and observed for 2 hours after replating into a new plate. Original magnifications: (A), (B), and (D), × 200; (C), × 100. (E through H) Giemsa staining was performed after cytocentrifuging nonadherent cells or aggregate cells (from D) from murine Lin⁻c-kit⁺ HPCs cultured for 14 days in the absence (E and G) or in the presence of mouse TNFα (F and H) combined with GM-CSF and SCF. Original magnifications: (E) and (F), × 160; (G) and (H), × 400.