Enhanced migration of healthy donor PBMC toward conditioned supernatants from alternative DC. “Wash out” supernatants collected 24 h after seeding mature DC in the absence of exogenous stimuli were used as a source of chemoattractants for immune cells. Total PBMC from healthy donors were loaded in a trans-well insert (3 μm pores) and allowed to migrate toward medium or DC “wash out” supernatants for 3 h. Migrated cells were analysed by flow cytometry to determine their total number (a) as well as their distribution into the different immune cell subpopulations (b) by using the indicated gating strategy (c). Data from seven different experiments are reported as mean ± SE after normalization to supernatants from gold standard DC. *p < 0.05 in the Tukey post hoc test.