Figure 7. CD34^–CD123⁺CD117^dimCD45RA⁺ cells preferentially give rise to CD5⁺ DCs.

(A) Gating strategy for cord blood progenitors. Cells were sorted as Lin^–DC^–CD10^–CD123^– (see also Supplemental Figure 3): CD34⁺CD117⁺ (blue); CD34^–CD117⁺ (orange); or Lin^–DC^–CD10^–CD123⁺CD34^–CD117^dimCD45RA⁺ (red). Histograms show the expression of CD5 and CD45RA on the different progenitor subsets. Expression on lineage⁺ cells is shown as a control (gray). (B) Sorted cord blood CD34⁺CD123^–CD117⁺, CD34⁻CD123^–CD117^–, or CD34⁻CD123⁺CD117^dim progenitors were cultured in the presence of GM-CSF, SCF, FLT3-L, and LTα/β. Flow cytometry plots, gated on live CD45⁺HLA-DR⁺CD11c⁺ cells, show culture output of CD1a⁺CD5⁺ DCs on day 7. Representative results of 4 independent experiments are shown. (C) Gating strategy for dermal progenitors. Cells were sorted as Lin^–DC^–CD10^– that were CD34⁺CD123^–CD117^– (blue); CD34^–CD123^–CD117⁺ (orange); or CD34⁻CD123⁺CD117^dim (red). Histograms show the expression of CD5 and CD45RA on the different progenitor subsets. (D) CD34⁺CD123^–CD117^–, CD34⁻CD123^–CD117⁺, or CD34⁻CD123⁺CD117^dim were isolated from human dermis. Flow cytometry plots are gated on live CD45⁺HLA-DR⁺CD11c⁺ cells and show the expression of HLA-DR⁺ or lineage⁺ cells that were differentiated from the different progenitors. (E) The plots show the expression of CD1c and CD5 on HLA-DR⁺ cells that differentiated from CD34⁻CD123⁺CD117^dim cells for 7 days in the presence of in the presence of GM-CSF (GM), SCF, and FLT3-L (FL) and either TNF-α or LTα/β. Representative results of 3 independent experiments are shown.