Generation and maturation of dendritic cells for clinical application under serum-free conditions

J Immunother. 2005 Nov-Dec;28(6):599-609. doi: 10.1097/01.cji.0000175491.21099.04.


Monocyte-derived dendritic cells (MoDCs) in clinical use for cancer immunotherapy are ideally generated in serum-free medium (SFM) with inclusion of a suitable maturation factor toward the end of the incubation period. Three good manfacturing practice (GMP) grade SFMs (AIM-V, X-VIVO 15, and X-VIVO 20) were compared with RPMI-1640, supplemented with 10% fetal bovine serum or 10% human serum. DCs generated for 7 days in SFM were less mature and secreted less interleukin (IL) 12p70 and IL-10 than DCs generated in 10% serum. DC yield was comparable in SFMs, and a greater proportion of cells was viable after maturation. Toll-like receptor (TLR) ligands were compared for their ability to induce cytokine secretion under serum-free conditions in the presence of interferon (IFN) gamma. With the exception of Poly I:C, TLR ligands stimulated high levels of IL-10 secretion. High levels of IL-12p70 were induced by two TLR4-mediated stimuli, lipopolysaccharide and Ribomunyl, a clinical-grade bacterial extract. When T-cell responses were compared in allogeneic mixed leukocyte reaction, DCs stimulated with Ribomunyl induced higher levels of IFNgamma than DCs stimulated with the cytokine cocktail: tumor necrosis factor-alpha, IL-1beta, IL-6, and prostaglandin E2. In the presence of IL-10 neutralizing antibodies, DC IL-12p70 production and T-cell IFNgamma were increased in vitro. Similarly, DCs stimulated with Ribomunyl, IFNgamma, and anti-IL-10 induced high levels of tetanus toxoid-specific T-cell proliferation and IFNgamma secretion. Thus, MoDCs generated in SFM efficiently stimulate T-cell IFNgamma production after maturation in the presence of a clinical-grade TLR4 agonist and IL-10 neutralization.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Cancer Vaccines
  • Cattle
  • Cell Differentiation
  • Cells, Cultured
  • Culture Media, Serum-Free*
  • Cytokines / biosynthesis*
  • Dendritic Cells / cytology*
  • Dendritic Cells / immunology
  • Humans
  • Monocytes / cytology
  • Serum
  • T-Lymphocytes / immunology
  • Toll-Like Receptor 4 / metabolism


  • Cancer Vaccines
  • Culture Media, Serum-Free
  • Cytokines
  • TLR4 protein, human
  • Toll-Like Receptor 4