Uptake of 2-hydroxyglutarate (HG) by dendritic cells (DCs) has no impact on DC viability and maturation. (a) Human DCs were differentiated from elutration-separated monocytes in the presence of FCS, GM-CSF, and IL-4. On day seven, immature DCs were stimulated with 100 ng/mL LPS in the presence or absence of 10 mM 2-HG for 24 h and viability was analyzed using the CASY cell analyzer system. Data represent the median of 10 independent experiments with DCs from different donors. (b) Uptake of D-2-HG and L-2-HG by DC was analyzed after stimulation with 100 ng/mL LPS in the presence or absence of 10 mM 2-HG for 1 h or 24 h. Cells were washed with PBS and intracellular levels of 2-HG were analyzed by mass spectrometry. Statistical analysis was performed with the Wilcoxon-Test (** p ≤ 0.01). (c,d) The expression of typical DC markers on DCs stimulated for 24 h with 100 ng/mL LPS in the absence or presence of 2-HG was analyzed by flow cytometry. (e) As a control, DCs were cultured in the absence or presence of 2-HG without LPS stimulation.