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CD34-derived cDC1s and cDC2s can be generated from blood progenitors. CD34-positive cells isolated from blood were differentiated into cDC1s and cDC2s in a 17-day protocol in which progenitors initially expand and afterward undergo skewing towards cDC1 and cDC2 differentiation. (A) Schematic representation of the 17-day protocol for DC generation. (B) Bar graph showing the percentage of cDC1s and cDC2s ± SEM at the end of the culture on day 17 ( n = 10). (C) Representative dot plot showing the presence of cDC1s and cDC2s based on the expression of CD141/CLEC9A and CD1c, respectively, after gating on viable single cells. (D) Violin plot showing the absolute number of cDC1s and cDC2s obtained with the current protocol, depending on the initial amount of cultured CD34-positive cells. Each dot represents the value of expanded cDC1s and cDC2s obtained from one independent donor ( n = 12). (E) Bar graphs show the percentage of positive DCs ± SEM for the given marker ( n = 3). CD34-derived cDC1s and cDC2s were characterized by flow cytometry to determine their lineage marker expression.

Journal: Oncoimmunology

Article Title: Ex vivo-generated conventional dendritic cells type 1 and type 2 from blood progenitors induce potent antigen-specific T-cell immunity

doi: 10.1080/2162402X.2026.2695692

Figure Lengend Snippet: CD34-derived cDC1s and cDC2s can be generated from blood progenitors. CD34-positive cells isolated from blood were differentiated into cDC1s and cDC2s in a 17-day protocol in which progenitors initially expand and afterward undergo skewing towards cDC1 and cDC2 differentiation. (A) Schematic representation of the 17-day protocol for DC generation. (B) Bar graph showing the percentage of cDC1s and cDC2s ± SEM at the end of the culture on day 17 ( n = 10). (C) Representative dot plot showing the presence of cDC1s and cDC2s based on the expression of CD141/CLEC9A and CD1c, respectively, after gating on viable single cells. (D) Violin plot showing the absolute number of cDC1s and cDC2s obtained with the current protocol, depending on the initial amount of cultured CD34-positive cells. Each dot represents the value of expanded cDC1s and cDC2s obtained from one independent donor ( n = 12). (E) Bar graphs show the percentage of positive DCs ± SEM for the given marker ( n = 3). CD34-derived cDC1s and cDC2s were characterized by flow cytometry to determine their lineage marker expression.

Article Snippet: Alternatively, DCs were enriched by MACS using CD141-Microbeads (130-090-512, Miltenyi Biotec) and CD1c Isolation kit (130-119-475, Miltenyi Biotec) sequentially (see supplementary figure 1C).

Techniques: Derivative Assay, Generated, Isolation, Expressing, Cell Culture, Marker, Flow Cytometry