Dendritic cells generated in roller bottle cultures have potent allo-stimulatory capability. Dendritic cells were generated in 850 cm2 roller bottles or T-175 static flasks as before using ITIP maturation. The floating cells were harvested, irradiated at 20 Gy and mixed with 50,000 allogeneic T-cell-enriched PBMC (plastic non-adherent PBMC) at different stimulator to responder ratios in 96-well plates. After 5 days, 1 μCi/well of 3H-thymidine was added to each well and the plates harvested 18 h later. Irradiated PBMC from the original DC donor were also used as stimulators as a control. The average cpm and standard deviation of triplicate cultures are shown for each stimulator type.