Efficient generation of pure CD103+ cDC1 in iDC1 cultures requires the +32 kb Irf8 enhancer (IMAGE)
Caption
(a-e) Bone marrow cells were cultured with FLT3L and GM-CSF (iCD103-DC) or with FLT3L, GM-CSF, and KitL (iDC1), followed by serial replating with FLT3L and GM-CSF every 7 days. Cells were analyzed by flow cytometry at the indicated time points. (a-c) One representative experiment including all three time points is shown. Similar results were obtained in four additional independent experiments, including one experiment containing day 23 and day 30 analyses and three additional experiments containing day 16 analyses. (a) Experimental scheme; (b) Frequencies and (c) total numbers of CD103+ cDC1 at the indicated time points; (d, e) One representative experiment of three independent experiments, each including four to five biological replicates, is shown; (d) Representative contour plot of CD103 versus CD172a expression among CD11c+ CD45R/B220- CD64- cDC from day 23 iDC1 cultures; (e) Expression of indicated markers in cDC1 and cDC2 from day 23 iDC1 cultures; (f-k) iDC1 cultures were generated from WT or Irf8+32-/- bone marrow and analyzed on day 23; (f-j) One representative of two independent experiments, each including five biological replicates per genotype, is shown. (f) Representative contour plots showing cDC1 and cDC2 populations among CD11c+ CD45R/B220- CD64- cDC; (g) Frequency and (h) total number of cDC1; (i) Frequency and (j) total number of cDC2; (k) Representative micrographs from one of six independent experiments (scale bars, 100 µm (top) and 20 µm (bottom)). ** p < 0.01, *** p < 0.001, **** p < 0.0001, NS = not statistically significant (unpaired two-tailed Student’s t-test). cDC1: conventional type 1 dendritic cells; iDC1: induced cDC1; FLT3L: fms-related tyrosine kinase 3 ligand; KitL: kit ligand; GM-CSF: granulocyte-macrophage colony-stimulating factor; WT: wild-type; APC: antigen-presenting cell; FACS: fluorescence activated cell sorting; cDC2: conventional type 2 dendritic cells.
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© Christian T. Mayer, et al. 2026. This is an Open Access article licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, sharing, adaptation, distribution and reproduction in any medium or format, for any purpose, even commercially, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
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