Functional characterization of monocyte-derived dendritic cells generated under serumfree culture conditions.
The culture of human monocyte-derived dendritic cells (DCs) is typically performed in media containing human or fetal calf serum, supplements with the potential to influence the cells phenotype and their functional properties. Published clinical trails based on serumfree cultured DCs reported the use of the commercially available medium AIMV. In this study, we directly compared DCs generated in AIMV medium ("AIMV/sf-DCs") with DCs generated in RPMI supplemented with 2% human serum ("RPMI/HS-DCs") in functional assays of potential relevance for vaccine application. Using TNF-alpha/PGE(2)/IL-1beta/IL-6 as maturation stimulus, AIMV/sf-DCs revealed to be comparable with RPMI/HS-DCs with regard to phenotypic expression of maturation markers, survival in vitro, migratory capacity and stimulation of lymphocyte proliferation except for CD1a which was expressed on a fraction of DCs only when cultured in serumfree AIMV medium. However, IL-12p70 production in response to Toll-like receptor (TLR) stimulating agents plus IFN-gamma was consistently lower in AIMV medium although also under serumfree culture conditions, nanogram quantities of IL-12 were produced. Together, DCs with functional characteristics important for in vivo application can be generated under defined serumfree conditions; however, medium and/or serum conditions appear to have strong influence on the production of relevant T cell differentiating cytokines.
Lehner
Manfred
M
Department of Cellular Therapy, Children's University Hospital, Erlangen, Germany.
Morhart
Patrick
P
Stilper
Andrea
A
Holter
Wolfgang
W
D018949:Antigens, CD1 / Q000032:analysis; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D016207:Cytokines / Q000096:biosynthesis / Q000494:pharmacology; D003713:Dendritic Cells / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D006801:Humans; D018664:Interleukin-12 / Q000096:biosynthesis; D008070:Lipopolysaccharides; D008562:Membrane Glycoproteins / Q000819:agonists; D010641:Phenotype; D021122:Protein Subunits / Q000096:biosynthesis; D011956:Receptors, Cell Surface / Q000819:agonists; D051193:Toll-Like Receptors
D018949:Antigens, CD1; C419393:CD1a antigen; D016895:Culture Media, Serum-Free; D016207:Cytokines; D008070:Lipopolysaccharides; D008562:Membrane Glycoproteins; D021122:Protein Subunits; D011956:Receptors, Cell Surface; D051193:Toll-Like Receptors; C482199:lipopolysaccharide, E coli O55-B5; D018664:Interleukin-12