Monocyte-derived DCs obtained after 4 days of maturation with TNF-α express the morphology and phenotype of mature DCs.

(A) Immature DCs cultured for 8 days in IL-4 and GM-CSF (left) or mature DCs cultured for 4 days in IL-4 and GM-CSF plus 4 days in IL-4 and GM-CSF and TNF-α (center and right) were cytospun onto glass slides and stained by Wright-Giemsa. Magnification  × 100. (B) (C) Flow cytometric analysis of immature DCs cultured for 8 days in IL-4 and GM-CSF (B) and mature DCs cultured for 4 days in IL-4 and GM-CSF plus 4 days in IL-4 and GM-CSF and TNF-α (C). DCs were double-stained with a panel of PE-conjugated mAbs and with FITC-conjugated mAb HLA-DR. Cells were also stained with isotype controls for the PE-conjugated mAbs and were double-stained with FITC-IgG2a, the isotype control for FITC-conjugated HLA-DR.