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

• Main Authors: Obermaier Bianca, Dauer Marc, Herten Jan, Schad Katharina, Endres Stefan, Eigler Andreas
• Format: Article
• Language: English
• Published: BMC 2003-01-01
• Series: Biological Procedures Online
• Subjects: Monocytes; Dendritic cells; Antigen presenting cells; Vaccination; Cell differentiation
• Online Access: http://www.biologicalprocedures.com/bpo/arts/1/62/m62.htm
• ISSN: 1480-9222

<p>We developed a new 2-day protocol for the generation of dendritic cells (DCs) from human monocytes <it>in vitro</it>. 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.