Figure 4.

Generation of human monocyte-derived DCs (moDCs) from PBMCs and the detection of DC activation markers after co-incubation with treated cancer cells. (a) PBMCs were isolated from buffy coat and seeded into an IgG pre-coated cell culture dish. On day 6 after differentiation, moDCs were co-cultured with differently treated MCF-7 breast cancer cells. After 24 h and 48 h of co-incubation, the activation markers of the moDCs were analysed using multicolor flow cytometry. The gating strategies for flow cytometry are shown (b–h). (b) After pre-gating on the singlets, the viable cells were detected (c,d). Then, gating on CD11c positive cells identified moDCs (e). Dot plots of CD83 (f), CD70 (g) and CD80 (h) expression on the cell surface of moDCs are exemplarily presented.