Open Access

Development of a new protocol for 2-day generation of mature dendritic cells from human monocytes

  • Bianca Obermaier1,
  • Marc Dauer2,
  • Jan Herten1,
  • Katharina Schad1,
  • Stefan Endres1 and
  • Andreas Eigler3Email author
Biological Procedures Online5:51197

Received: 18 August 2003

Accepted: 22 September 2003


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 terms

MonocytesDendritic cellsAntigen presenting cellsVaccinationCell differentiation