Generation of large numbers of dendritic cells in a closed system using Cell Factories

J Immunol Methods. 2002 Jun 1;264(1-2):135-51. doi: 10.1016/s0022-1759(02)00099-6.


There is a growing interest in using dendritic cells (DC) for vaccine approaches in the treatment of cancer and infectious diseases. This requires a reproducible method for the generation of large numbers of DC in a closed culture system suitable for clinical use and conforming to the current guidelines of good manufacturing practices. We designed a system in which the DC were generated in a closed system from adherent monocytes using Cell Factories (DC-CF). Monocytes were enriched from apheresis products by adherence and then cultured in the presence of AB serum or autologous plasma and GM-CSF and IL-4 for 6 days. The DC generated in Cell Factories were extensively compared to research-grade DC generated in conventional tissue culture flasks (DC-TCF). At day 6, the immature DC were harvested and the yield, the viability, the immunophenotype and the functional characteristics of the DC were compared.DC-CF and DC-TCF showed similar viability and purity and scored equally when tested for stability, dextran and latex bead uptake, in MLR and in the activation of influenza-specific memory cells after electroporation with influenza matrix protein 1 (IMP1) mRNA. These data indicated that large numbers of functional clinical-grade DC could be generated from adherent cells in a closed system using Cell Factories.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Adhesion / immunology
  • Cell Count
  • Cell Culture Techniques / instrumentation*
  • Cell Culture Techniques / methods*
  • Cell Differentiation / immunology
  • Cell Line
  • Clone Cells
  • Cryopreservation / methods
  • Culture Techniques / instrumentation
  • Dendritic Cells / cytology*
  • Dendritic Cells / immunology
  • Electroporation / methods
  • Humans
  • Immunologic Memory / genetics
  • Lymphocyte Activation / genetics
  • Mice
  • Monocytes / cytology
  • RNA, Messenger / metabolism
  • T-Lymphocyte Subsets / immunology
  • T-Lymphocyte Subsets / metabolism
  • T-Lymphocyte Subsets / virology
  • Viral Matrix Proteins / genetics
  • Viral Matrix Proteins / immunology
  • Viral Matrix Proteins / metabolism


  • M-protein, influenza virus
  • RNA, Messenger
  • Viral Matrix Proteins