Surface-mediated priming during in vitro generation of monocyte-derived dendritic cells.
Ex vivo-generated human dendritic cells (DC) are most commonly generated from monocytes using standard cell culture dishes. To elucidate the effect of the plastic surface during the differentiation process, we compared a standard adhesive plastic dish with four different mainly non-adherent surfaces. Untouched monocytes were cultured for 3 days in the presence of IL-4 and GM-CSF. Time-lapse videos were recorded, and the phenotype of the cells was analysed by flow cytometry. The cytokine profiles were analysed using a 25-plex cytokine assay. The use of non-adherent surfaces led to a significant reduction in expression of CD14 and CD38, and a significant increase in expression of CD86 compared to standard culture dishes. Expression levels of DC-SIGN and PD-L2 were reduced significantly on cells cultured on non-adherent surfaces. The cytokine production was independent on the surface used. The surface-mediated priming should therefore be considered when aiming to induce specific immune responses. This is especially important with regard to DC-based immunotherapy, where an adjustment of the surface during the DC generation process might have highly beneficial effects.
Department of Biomedicine, University of Bergen, Bergen, Norway.
A
A
Sauter
Y
Y
Mc Duffie
H
H
Boehm
A
A
Martinez
J P
JP
Spatz
S
S
Appel
D051997:ADP-ribosyl Cyclase 1 / Q000096:biosynthesis; D000328:Adult; D051940:B7-2 Antigen / Q000096:biosynthesis; D002448:Cell Adhesion / Q000502:physiology*; D015815:Cell Adhesion Molecules / Q000096:biosynthesis; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000276:immunology*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D015847:Interleukin-4 / Q000494:pharmacology; D037181:Lectins, C-Type / Q000096:biosynthesis; D018950:Lipopolysaccharide Receptors / Q000096:biosynthesis; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D009000:Monocytes / Q000276:immunology*; D010969:Plastics / Q000494:pharmacology; D061027:Programmed Cell Death 1 Ligand 2 Protein / Q000096:biosynthesis; D011956:Receptors, Cell Surface / Q000096:biosynthesis; D013499:Surface Properties; D055815:Young Adult; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
D051997:ADP-ribosyl Cyclase 1; D051940:B7-2 Antigen; D015815:Cell Adhesion Molecules; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D015847:Interleukin-4; D037181:Lectins, C-Type; D018950:Lipopolysaccharide Receptors; D008562:Membrane Glycoproteins; D010969:Plastics; D061027:Programmed Cell Death 1 Ligand 2 Protein; D011956:Receptors, Cell Surface; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin; C497964:CD86 protein, human; C508598:IL4 protein, human; C556977:PDCD1LG2 protein, human; C498154:CD38 protein, human