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To remove the hematopoietic cells and to prevent overgrowth of the cultures with macrophages, the bone marrow suspension was depleted from CD45+ cells by negative immuno magnetic cell selection using the AutoMACS device (Miltenyi Biotec, Bergisch Gladbach, Germany). Briefly, bone marrow cells (BMC) were resuspended in MACS buffer (PBS + 1% FCS + 2 mM EDTA) and stained for 30 min nike roshe run donker grijs on ice with cychrome labelled rat anti mouse CD45 (BD Pharmingen, Alphen a/d Rijn, The Netherlands). Following labelling cells were washed with MACS buffer and resuspended in MACS buffer at a concentration of 107cells/40l, after which the BMC were incubated for 20 min at 4 with 10 l of paramagnetic beads (goat anti rat Ig microbeads; Miltenyi Biotec) per 107 cells.
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