Development of a standardized protocol for reproducible generation of matured monocyte-derived dendritic cells suitable for clinical application |
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Authors: | HR Bohnenkamp T Noll |
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Institution: | (1) Research Center Juelich GmbH, Institute of Biotechnology 2, 52428 Juelich, Germany |
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Abstract: | There is increasing interest in the generation of dendritic cells (DC) for cancer immunotherapy. In order to utilize DC in
clinical trials it is necessary to have standardized, reproducible and easy to use protocols. We describe here the process
development for the generation of DC as the result of investigation of culture conditions as well as consumption rates of
medium and cytokines. Our studies demonstrate that highly viable DC (93 ± 2%) can be produced from CD14+ enriched monocytes via immunomagnetic beads in a high yield (31 ± 6%) with X-VIVO 15, 400 U ml−1 GM-CSF and 2000 U ml−1 IL-4 without serum and feeding. For the maturation of DC different cocktails (TNF-α, IL-1β, IL-6, PGE2 and TNF-α, PGE2) were compared. In both cases cells expressed typical surface molecules of mature DC and induced high proliferative responses
in mixed lymphocyte reactions which led to IFN-γ producing T-lymphocytes. The data suggest that the use of this optimized,
easy to use protocol results in highly mature DC.
This revised version was published online in July 2006 with corrections to the Cover Date. |
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Keywords: | Dendritic cells Immunotherapy Monocyte enrichment Serum-free |
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