About Dendritic CellsVIEW PRODUCT LISTING
We generate our myeloid dendritic cells (DC) by enriching monocytes, then culturing them in the presence of IL-4 and GM-CSF. Cells are collected after 5–7 days of culture and characterized by flow cytometric analysis for expression of CD11c, CD14, CD86, HLA-DR, CD1a, CD3 and CD19. The cells are frozen as immature DC but can be stimulated to full maturation as shown in the figure below.
We also have a variety of mouse dendritic cells available.
Dendritic Cell Maturation Using Poly I:C
Directly after thawing, we plated dendritic cells in a 24 well plate at 106 cells/2 mL/well, in X-VIVO 15 (immature DCs) or X-VIVO 15 + 20 μg/mL poly I:C (mature DCs) for 48 hours in a 37°C, 6% CO2 humidified incubator.
We harvested and analyzed DCs for the surface expression of CD40, CD86, CD54, CD80, CD83 and HLA-DR/-DP/-DQ on a FACScan (Becton Dickinson, San Jose, CA) flow cytometer. Histograms representing binding of antigen-specific fluorescent-labeled antibody to immature DCs (black solid line) or mature DCs (dark red histogram), as well as binding of isotype controls (dashed lines) are shown for each surface antigen tested (see figure below).
This data shows an increase of expression of each of the surface antigens tested, demonstrating DC maturation after 48 hours of culture in the presence of poly I:C.