A functional comparison of mature human dendritic cells prepared in fluorinated ethylene-propylene bags or polystyrene flasks.
BACKGROUND
Fluorinated ethylene-propylene (FEP) bags have been used instead of polystyrene (PS) flasks for ex vivo clinical-scale production of human dendritic cells (DCs) to facilitate closed-system recovery of these highly adherent cells. To assess the impact of DC culture on this nonadherent surface, the function of DCs generated in FEP and PS was compared.


METHODS
Cell yield, phenotype, cytokine production, migration, and antigen-presenting activity were measured in DCs prepared from peripheral blood monocytes in FEP bags or PS flasks with medium supplemented with serum, interleukin (IL)-4, and granulocyte-macrophage-colony-stimulating factor for 5 days to induce DC differentiation and CD40L or poly(I:C) plus interferon-gamma to promote maturation.


RESULTS
DCs cultured in FEP or PS had comparable cell yield, viability, and CD83 and CCR7 expression. DCs generated in FEP, however, produced significantly less IL-12 and IL-10 during maturation, and differences persisted on rechallenge after harvest. FEP-cultured DCs migrated spontaneously or in response to CCR7 ligand more actively than PS-cultured DCs, but this difference was not significant. Mature DCs prepared in FEP and PS were equipotent in stimulating peptide-specific CD8 T-cell expansion in vitro.


CONCLUSIONS
FEP- and PS-cultured DCs are similar in phenotype and in some functional measures, but FEP markedly reduces DC production of IL-12 and IL-10. This phenomenon presumably reflects intracellular changes linked to the absence of a surface for firm cell adherence. Given the importance of these cytokines in the immune response, these changes could have a significant impact on DC function in vivo.
Kurlander
Roger J
RJ
Department of Laboratory Medicine, NIH Clinical Center, National Institutes of Health, Bethesda, Maryland 20892-1508, USA. rkurlander@nih.gov
Tawab
Abdul
A
Fan
Yong
Y
Carter
Charles S
CS
Read
Elizabeth J
EJ
D000938:Antigen-Presenting Cells / Q000187:drug effects / Q000276:immunology; D023201:CD40 Ligand / Q000494:pharmacology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002452:Cell Count; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000166:cytology* / Q000187:drug effects / Q000276:immunology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D016753:Interleukin-10 / Q000096:biosynthesis; D018664:Interleukin-12 / Q000096:biosynthesis; D015847:Interleukin-4 / Q000494:pharmacology; D009000:Monocytes / Q000166:cytology; D010455:Peptides / Q000494:pharmacology; D011137:Polystyrenes / Q000494:pharmacology*; D011138:Polytetrafluoroethylene / Q000031:analogs & derivatives* / Q000494:pharmacology; D013997:Time Factors; D005465:Fluorocarbon Polymers
D023201:CD40 Ligand; D003470:Culture Media; D016207:Cytokines; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D007371:Interferon-gamma; D016753:Interleukin-10; D018664:Interleukin-12; D015847:Interleukin-4; D010455:Peptides; D011137:Polystyrenes; D011138:Polytetrafluoroethylene; C096305:fluorinated ethylene propylene; D005465:Fluorocarbon Polymers