Journal
JOURNAL OF IMMUNOLOGICAL METHODS
Volume 294, Issue 1-2, Pages 67-80Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jim.2004.08.010
Keywords
monocytes; dendritic cells; apoptosis; annexin V; yield
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Monocyte-derived dendritic cells (DC) are currently under extensive evaluation as cell vaccines for cancer treatment. The requirement for large-scale cell products demands optimized and standardized protocols. However, the yield of DCs from inoculated monocytes is reported to be always lower than 50%. In this present study we investigated whether this cell loss was caused 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/propidium iodide assay. We found 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. Monocytes were differentiated with GM-CSF and interleukin-4 into a viable, non-apoptotic population of immature dendritic cells. Addition of tumor necrosis factor-alpha and prostaglandin E-2 resulted in fully matured dendritic cells, which were evaluated by phenotypic analysis and by allogeneic and MHC class-I-restricted T-cell responses. About 90.2 +/- 16.7% of the non-apoptotic monocyte population differentiated to viable matured dendritic cells. These results indicate that the yield of dendritic cells is mainly influenced by the percentage of apoptotic cells in the inoculum, and this has implications for DC generation in clinical applications. (C) 2004 Elsevier B.V. All rights reserved.
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