Dendritic cell isolation and flow cytometry analysis for myeloid and lymphoid DC.

(A) Isolation scheme for low-density MHC-II⁺ cells. Low-density cells are isolated by density gradient centrifugation followed by magnetic bead enrichment for MHC-II⁺ cells. Cells are subjected to successive rounds of positive selection until more than 95% are MHC II⁺ cells, as determined by flow cytometry. (B) Low-density MHC-II⁺ cells (panel 1) are then stained for DC markers CD11c (panel 2), CD8α (panel 3), or DEC-205 (last panels) and analyzed by flow cytometry. Representative staining histograms are presented with isotype controls provided. Dead cells were excluded from analysis by propidium iodide exclusion and scattergating for size.