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Development of a new protocol for 2-day generation of mature dendritic cells from human monocytes
Biological Procedures Online volume 5, pages 197–203 (2003)
Abstract
We developed a new 2-day protocol for the generation of dendritic cells (DCs) from human monocytesin vitro. First, we demonstrated that 24 hours of culture with GM-CSF and IL-4 are sufficient to generate immature DCs capable of antigen uptake. We then compared two different strategies for DC maturation: proinflammatory mediators were either added together with GM-CSF and IL-4 from the beginning of cell culture or added after 24 hours of differentiation with GM-CSF and IL-4. After 48 hours of total culture period, expression of activation markers was more pronounced in cells generated by the 2-step differentiation and activation method. Our new protocol for 2-day DC differentiation reduces labor, cost and time and also reliably renders high numbers of mature and viable DCs.
Abbreviations
- DC:
-
dendritic cell
- moDC:
-
monocyte-derived DC
- PBMC:
-
peripheral blood mononuclear cells
- CD40L:
-
soluble CD40 ligand-trimer
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B.O. and M.D. contributed equally to this manuscript.
Published: October 24, 2003
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Obermaier, B., Dauer, M., Herten, J. et al. Development of a new protocol for 2-day generation of mature dendritic cells from human monocytes. Biol. Proced. Online 5, 197–203 (2003). https://doi.org/10.1251/bpo62
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DOI: https://doi.org/10.1251/bpo62