1. In vitro generation of murine myeloid dendritic cells from CD34-positive precursors
- Author
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Stephen, Tom-Li, Harms, Fabian, Fabri, Mario, Flenner, Eva, Bessler, Martina, Hafke, Helena, Meemboor, Sonja, Kalka, Christoph, and Kalka-Moll, Wiltrud
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DENDRITIC cells , *MYELOID leukemia , *GRANULOCYTE-macrophage colony-stimulating factor , *LABORATORY mice , *ANTIGEN presenting cells , *BONE marrow cells - Abstract
Abstract: Dendritic cells (DCs) link the innate and adaptive immune system. Currently, murine DCs for cell biology investigations are developed from MHC class II-negative bone marrow (BM) precursor cells, non-depleted BM cells or BM monocytes in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF). Here we demonstrate an isolation procedure of functionally intact myeloid CD11c+ CD11b+ DCs derived from murine CD34-positive precursors. DCs derived from CD34+ cells show functional internalization, maturation, cytokine secretion, MHC-restricted antigen presentation, and MHCII retrograde transport of antigens from the lysosomes to the cell surface. In comparison to the established method, the advantages of this isolation procedure are a shorter cultivation period, a superior transfection efficiency, the yield of a purer and more homogeneous population of immature DCs, and less consumption of cell culture medium and GM-CSF. The new isolation procedure and the functional quality of CD34+ cell-derived murine myeloid DCs make them ideally suited for immunology and cell biology studies. [Copyright &y& Elsevier]
- Published
- 2009
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