Monocyte-derived dendritic cell (MoDC) differentiation

MoDCs were differentiated from monocytes isolated from PBMCs (via their ability to bind to plastic surfaces) and were cultivated in RPMI supplemented with 5% FCS (Biochrom), 1% penicillin-streptomycin (Gibco), 1% l-glutamine (Gibco) 1 mM HEPES and 1 mM sodium pyruvate (Sigma) in the presence of 800 U/ml GM-CSF (Peprotech) and 500 U/ml IL-4 (Peprotech). Cells were fed on day 3 and day 5. Experiments were performed on day 6 by co-incubation of MoDCs with heat-aggregated ICs after pre-conditioning of MoDCs with 0.5 ng/ml LPS.