Generation of large numbers of dendritic cells in a closed system using Cell Factories.
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.
Laboratory of Molecular and Cellular Therapy, Department of Physiology-Immunology, Medical School of the Vrije Universiteit Brussel (VUB), Laarbeeklaan 103/E, Brussels, Belgium.
Sandra
S
Tuyaerts
Sofie M
SM
Noppe
Jurgen
J
Corthals
Karine
K
Breckpot
Carlo
C
Heirman
Catherine
C
De Greef
Ivan
I
Van Riet
Kris
K
Thielemans
D000818:Animals; D002448:Cell Adhesion / Q000276:immunology; D002452:Cell Count; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002454:Cell Differentiation / Q000276:immunology; D002460:Cell Line; D002999:Clone Cells; D015925:Cryopreservation / Q000379:methods; D046508:Culture Techniques / Q000295:instrumentation; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology; D018274:Electroporation / Q000379:methods; D006801:Humans; D007156:Immunologic Memory / Q000235:genetics; D008213:Lymphocyte Activation / Q000235:genetics; D051379:Mice; D009000:Monocytes / Q000166:cytology; D012333:RNA, Messenger / Q000378:metabolism; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism / Q000821:virology; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
C031540:M-protein, influenza virus; D012333:RNA, Messenger; D014763:Viral Matrix Proteins