Recent reports have described a new strategy for differentiation and maturation of monocyte (Mo)-derived dendritic cells (DC) within only 48-72 h of in vitro culture (fast-DC). Mature fast-DC are as effective as mature standard-DC (generated in 7-10 days of in vitro culture) in priming and propagation of antigen-specific T-cell responses. The use of fast-DC not only reduces labor and supply cost, as well as workload and time, but also increases the DC yield from Mo, which may facilitate DC-based immunotherapy for cancer patients. Detailed protocols for generation, pulsing with different antigen sources, and transduction with adenoviral vector of Mo-derived mature fast-DC as well as using of fast-DC for priming and propagation of antigen-specific cytotoxic T-cell effectors will be described here.