Book	FZJ-2016-03840

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Bioprocess Development for the Generation of Monocyte-derived Dendritic Cells: applicability in breast cancer immunotherapy

Bohnenkamp, H. R. (Corresponding author)FZJ*

2004
Forschungszentrum Jülich GmbH Zentralbibliothek, Verlag Jülich
ISBN: 3-89336-364-5

Please use a persistent id in citations: http://hdl.handle.net/2128/11873

Abstract: Monocyte-derived dendritic cells (DCs) are currently under extensive evaluation as cell vaccines for cancer treatment. The requirement for large-scale cell products demands optimized and standardized protocols. This study demonstrates that highly viable DCs (93.4 ± 5.9%) can be produced from CD14$^{+}$ monocytes enriched via immunomagnetic beads in a high yield (66.3 ± 13.6%) and purity (95.4 ± 2.1%) with X-VIVO 15, 400$\frac{U}{ml}$ GM-CSF and 2000$\frac{U}{mL}$ IL-4 without serum and feeding. For the maturation of DCs different cytokine combinations (TNF-$\alpha$, IL-1$\beta$, IL-6, PGE$_{2}$ and TNF-$\alpha$, PGE$_{2}$) were compared. In both cases cells expressed typical surface molecules of mature DCs and induced high proliferative responses in mixed leukocyte reactions which led to IFN-$_{\gamma}$ producing T lymphocytes. Comparison of dendritic cells from blood from healthy donors with breast cancer patients’ blood demonstrated no differences regarding yield or quality of fully matured dendritic cells. Furthermore, this study aimed to investigate if the yield is determined by the properties of the starting population of inoculated monocytes. CD14$^{+}$ cells were enriched by immunomagnetic-bead selection and analyzed for apoptosis by an annexin V / propidiumiodide assay. It was demonstrated that 37.8±11.1% (n = 8) of freshly isolated monocytes from buffy coats of healthy donors underwent programmed cell death. Further analysis of the fate of apoptotic cells during differentiation suggested phagocytosis. The yield of viable matured dendritic cells from non-apoptotic monocytes was calculated to be 90.2±16.7%. These results indicate that the yield of dendritic cells is mainly influenced by the percentage of apoptotic cells in the inoculum, and impact on DC generation for clinical application. For cancer immunotherapy the loading of DCs with whole tumor cell lysate preparations represents a simple and promising approach to utilize all potential known and unknown [...]

Note: Universität Bonn, Diss., 2004

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Contributing Institute(s):

1. Biotechnologie 2 (IBT-2)

Research Program(s):

1. 899 - ohne Topic (POF3-899) (POF3-899)

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Database coverage:

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