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Biological Procedures Online

, Volume 5, Issue 1, pp 197–203

Received: 18 August 2003Revised: 21 September 2003Accepted: 22 September 2003DOI: 10.1251-bpo62

Cite this article as: Obermaier, B., Dauer, M., Herten, J. et al. Biol. Proced. Online 2003 5: 197. doi:10.1251-bpo62


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.

Indexing termsMonocytes Dendritic cells Antigen presenting cells Vaccination Cell differentiation AbbreviationsDCdendritic cell

moDCmonocyte-derived DC

PBMCperipheral blood mononuclear cells

CD40Lsoluble CD40 ligand-trimer

B.O. and M.D. contributed equally to this manuscript.

Published: October 24, 2003

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Author: Bianca Obermaier - Marc Dauer - Jan Herten - Katharina Schad - Stefan Endres - Andreas Eigler


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