In vitro generation of murine myeloid dendritic cells from CD34-positive precursors |
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Authors: | Tom-Li Stephen Fabian Harms Mario Fabri Martina Bessler Sonja Meemboor Wiltrud Kalka-Moll |
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Affiliation: | a Institute for Medical Microbiology, Immunology and Hygiene, Medical Center, University of Cologne, Goldenfelsstrasse 19-21, 50935 Cologne, Germany b Swiss Cardiovascular Center, Division of Vascular Medicine, Inselspital, Bern University Hospital, and University of Bern,Freiburgstrasse, 3010 Bern, Switzerland |
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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. |
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Keywords: | Murine dendritic cells CD34 Bone marrow Antigen presentation |
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