title: Comparison of media and serum supplementation for generation of monophosphoryl lipid A/interferon-γ-matured type I dendritic cells for immunotherapy.
abstract: OBJECTIVE
  Ex vivo-generated monocyte-derived dendritic cells (DCs) matured with monophosphoryl lipid A (MPLA) and interferon-γ (IFN-γ) can be used as cancer immunotherapy. MPLA/IFN-γ DCs induce Th1 T cell responses and have migratory capacity. Different culture regimens have been used for generation of immunotherapeutic DCs, with varying results. In the present study, culture conditions for MPLA/IFN-γ-matured type I DCs were optimized for clinical application.
  
  
  METHODS
  DCs were generated from monocytes in the clinical grade culture media CellGro DC, AIM V or X-VIVO 15 in the absence or presence of 2% human serum (HS) and matured with the use of MPLA/IFN-γ. DC yield and DC functionality were assessed. DC functionality was determined by means of analysis of cytokines in culture supernatant, migratory capacity, expression of co-stimulatory molecules, T cell stimulatory capacity of DCs and T helper cell (Th) polarization by the DCs.
  
  
  RESULTS
  DCs generated in the presence of 2% HS produced low amounts of pro-inflammatory cytokines and could not migrate irrespective of the medium used. In the absence of HS, MPLA/IFN-γ DCs generated in X-VIVO did not migrate either. MPLA/IFN-γ DCs generated in AIM V have slightly lower capacity to induce Th1 cells than do DCs generated in CellGro or X-VIVO.
  
  
  CONCLUSIONS
  Addition of HS to different GMP culture media is detrimental for pro-inflammatory DC maturation and migration. In the absence of serum, CellGro is the most optimal medium tested for generation of migratory and Th1-inducing MPLA/IFN-γ DCs for cancer immunotherapy.
authors: Department of Immunopathology, Sanquin Blood Supply, Division of Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors: Sonja T H M
authors: ST
authors: Kolanowski
authors: Department of Immunopathology, Sanquin Blood Supply, Division of Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors: Lathees
authors: L
authors: Sritharan
authors: Department of Immunopathology, Sanquin Blood Supply, Division of Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors: Suzanne N
authors: SN
authors: Lissenberg-Thunnissen
authors: Department of Immunopathology, Sanquin Blood Supply, Division of Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors: Gijs M W
authors: GM
authors: Van Schijndel
authors: Department of Immunopathology, Sanquin Blood Supply, Division of Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
authors: S Marieke
authors: SM
authors: Van Ham
authors: Department of Immunopathology, Sanquin Blood Supply, Division of Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands. Electronic address: a.tenbrinke@sanquin.nl.
authors: Anja
authors: A
authors: ten Brinke
mesh_terms: D002454:Cell Differentiation / Q000276:immunology; D049109:Cell Proliferation / Q000235:genetics; D064987:Cell- and Tissue-Based Therapy / Q000379:methods; D003470:Culture Media; D003713:Dendritic Cells / Q000166:cytology* / Q000637:transplantation; D006801:Humans; D007167:Immunotherapy / Q000379:methods; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D008050:Lipid A / Q000031:analogs & derivatives / Q000276:immunology / Q000378:metabolism; D009369:Neoplasms / Q000276:immunology / Q000628:therapy; D018417:Th1 Cells / Q000276:immunology
chemical_list: D003470:Culture Media; D008050:Lipid A; D007371:Interferon-gamma; C048436:monophosphoryl lipid A
keywords: clinical grade dendritic cells; culture media; immunotherapy; monophosphoryl lipid A; serum