cd14 microbeads miltenyi biotec  (Miltenyi Biotec)

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: Ex vivo-generated cDC1s and cDC2s acquire a matured state upon TLR challenge. Enriched cDC1s and cDC2s were overnight stimulated with a maturation cocktail containing R848 and Poly(IC), and the next day, their phenotype and cytokine/chemokine production were characterized. (A) Representative histograms showing the expression levels of CD83, CD40, and CCR7 upon maturation on DC2s (blue histograms) and cDC1s (red histograms). (B) Maturation marker expression on cDC2s (blue dots) and cDC1s (red dots) is displayed as either percentage of positive DCs (CD80, CD86, CD83, CD40, and CCR7) or relative geometric mean fluorescence intensity (gMFI) (HLA-DR). Relative gMFI expression was determined by normalizing to the average gMFI of untreated cDC1s or cDC2s. Scatter dot plots show the mean ± SEM with each data point representing an individual DC donor ( n ≥ 8). (C) Scatter dot plots show the mean ± SEM of the raw concentrations of the indicated cytokines and chemokines. Each data point represents an individual DC donor ( n ≥ 8). Statistical significance was calculated for paired data sets with a paired t-test or a Wilcoxon test, whereas for unpaired data sets significancy was calculated using a Mann–Whitney test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: After 2 h of coculture at 37oC, cells were stained to identify DC2s (CD1c-FITC, 1:25, 130-113-301, Miltenyi Biotec) and cDC1s (CD141 PerCP-Cy5.5, 1:25, 344112, BioLegend) prior to analysis using a BD FACSLyricTM.

Techniques: Ex Vivo, Generated, Expressing, Marker, Fluorescence, MANN-WHITNEY

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

Journal: The Journal of Cell Biology

Article Title: Galectin-9 regulates dendritic cell polarity and uropod contraction by modulating RhoA activity

doi: 10.1083/jcb.202404079

Figure Lengend Snippet: cDC2 cell purity and supplementary phenotype analysis. (A) Gating strategy to determine cDC2 purity after isolation from PBMCs. cDC2s were stained for CD1c, CD11c, CD14, and CD20. A first gate is set based on the physical FCS-A/SSC-A parameters followed by selecting on negative cells for CD14 and CD19. cDC2s are then identified as positive cells for CD11c and CD1c. (B) Surface galectin-9 and CCR7 expression in galectin-9 and CCR7-positive cDC2s. Graph shows average ± SEM gMFI of five individual donors. One-way ANOVA followed by Dunnett’s test for multiple comparisons was performed. *P < 0.05; **P < 0.01; ***P < 0.001. gMFI, geometric mean fluorescence intensity.

Article Snippet: Human cDC2s were isolated from peripheral blood mononuclear cells derived from healthy individuals (Sanquin, Nijmegen, The Netherlands) using the MACS CD1c + isolation kit (130-119-475; Miltenyi Biotec) according to the manufacturer’s instructions.

Techniques: Isolation, Staining, Expressing, Fluorescence