Figure 1.
Figure 1. Simultaneous isolation of pure populations of human neutrophils, monocytes, and DC subsets by flow cytometry cell sorting. (A) Pure populations of human CD66abce+ neutrophils, CD14+ and CD14−CD16+ monocytes, CD123+ PDCs, and CD11c+ MDC subsets (BDCA-1+ and BDCA-3+ subsets) were isolated from the elutriated monocyte-fraction after apheresis by flow cytometry sorting according to the depicted gating strategy. CD14−CD16+ monocytes are now indicated as CD16+ monocytes only for simplicity. (B) Representative flow histograms show the expression levels of the indicated markers on the sorted APC populations. (C) Sorted CD66abce+ neutrophils coexpress CD15. (D) Representative flow histograms show the live/dead staining (LIVE/DEAD fixable dead cell stain kit) on sorted neutrophils over time. Neutrophils showed high viability for at least 30 hours and can be maintained in culture longer. A representative donor of 6 is shown. FSC-A, forward scatter area; FSC-H, forward scatter height; SSC-A, side scatter area.

Simultaneous isolation of pure populations of human neutrophils, monocytes, and DC subsets by flow cytometry cell sorting. (A) Pure populations of human CD66abce+ neutrophils, CD14+ and CD14CD16+ monocytes, CD123+ PDCs, and CD11c+ MDC subsets (BDCA-1+ and BDCA-3+ subsets) were isolated from the elutriated monocyte-fraction after apheresis by flow cytometry sorting according to the depicted gating strategy. CD14CD16+ monocytes are now indicated as CD16+ monocytes only for simplicity. (B) Representative flow histograms show the expression levels of the indicated markers on the sorted APC populations. (C) Sorted CD66abce+ neutrophils coexpress CD15. (D) Representative flow histograms show the live/dead staining (LIVE/DEAD fixable dead cell stain kit) on sorted neutrophils over time. Neutrophils showed high viability for at least 30 hours and can be maintained in culture longer. A representative donor of 6 is shown. FSC-A, forward scatter area; FSC-H, forward scatter height; SSC-A, side scatter area.

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