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. Author manuscript; available in PMC: 2012 Apr 1.

Published in final edited form as: Immunobiology. 2010 Sep 24;216(4):447–456. doi: 10.1016/j.imbio.2010.09.008

(Panel A) Flow cytometry dot plot showing dendritic cells differentiated from monocytes isolated by ficoll gradient centrifugation from human umbilical cord blood. The CD14+ monocytes were supplemented with granulocyte macrophage colony stimulating factor (GMCSF) and IL-4 and were cultured for 6 days to obtain CD14-HLA-DR+CD11c+ immature human DCs. To determine the viability of DCs isolated from the entire population of collected cells (Upper left panel), 7-AAD negative cells were gated (Upper right panel) and analyzed for expression of the surface marker CD14 (Lower left panel). The CD14- cells were gated and analyzed for co-expression of HLA-DR and CD11C differentiation markers by flow cytometry. (Panel B) The histograms depict the expression of dendritic cell CD83 and CD86 after stimulation of DCs for 48 hours with CTB, CTB-INS, Insulin and PMA Ionomycin. Maturation state of the DCs was determined by flow cytometric analysis. The data are representative of three independent experiments with comparable results.

(Panel A) Flow cytometry dot plot showing dendritic cells differentiated from monocytes isolated by ficoll gradient centrifugation from human umbilical cord blood. The CD14+ monocytes were supplemented with granulocyte macrophage colony stimulating factor (GMCSF) and IL-4 and were cultured for 6 days to obtain CD14-HLA-DR+CD11c+ immature human DCs. To determine the viability of DCs isolated from the entire population of collected cells (Upper left panel), 7-AAD negative cells were gated (Upper right panel) and analyzed for expression of the surface marker CD14 (Lower left panel). The CD14- cells were gated and analyzed for co-expression of HLA-DR and CD11C differentiation markers by flow cytometry. (Panel B) The histograms depict the expression of dendritic cell CD83 and CD86 after stimulation of DCs for 48 hours with CTB, CTB-INS, Insulin and PMA Ionomycin. Maturation state of the DCs was determined by flow cytometric analysis. The data are representative of three independent experiments with comparable results.