Analysis of methods for the generation of dendritic cells from human peripheral blood monocytes

Dendritic cells (DC) are highly efficient antigen-presenting cells that ini tiate the primary immune response. Several laboratories have developed cult ure systems for human DC from peripheral blood monocytes. Most of these stu dies have used fetal calf serum (FCS) containing culture conditions that ar e inappropriate for human application. GM-CSF and IL-4 were Used to make im mature DC. The monocyte-conditioned medium (MCM) was used to induce the fin al maturation of DC; Using the previously described methods, the quality of MCM has unpredictable variations. Therefore using a defined cocktail of gr owth factors for the generation bf mature DC would be advantageous for expe rimental as well as clinical purposes. In this study, it is suggested that combinations of both GM-CSF/IL-4 or GM-CSF/IL-13 could be used as the first -step culture to produce immature DC, and that cytokine cocktail (GM-CSF, I L-4, IL-1 beta, TNF-alpha, IL-6, PGE(2)) was as efficient as MCM for the se cond step-culture to produce fully maturated DC. Here, we have generated an easily reproducible culture system for DC that allows for the generation o f large amounts of immature and mature DC, and we also now have established the method in a FCS-free system that is suitable for clinical use.