Development of a new protocol for 2-day generation of mature dendritic cells from human monocytes.
We developed a new 2-day protocol for the generation of dendritic cells (DCs) from human monocytes in 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.
Obermaier
Bianca
B
Division of Clinical Pharmacology, Medizinische Klinik Innenstadt, University of Munich. Ziemssenstr. 1, 80336 Munich. Germany. B.O. and M.D. contributed equally to this manuscript.
Dauer
Marc
M
Herten
Jan
J
Schad
Katharina
K
Endres
Stefan
S
Eigler
Andreas
A