Isolation and Differentiation of Macrophages

Human Serum and PBMC (peripheral blood mononuclear cells) were prepared as described previously (53 (link)). Monocytes were isolated through adherence of PBMC in monocyte attachment medium (PromoCell) for 30 min at 37°C. Monocytes were differentiated into macrophages in serum-free X-vivo medium (Lonza) containing 50 U/mL penicillin and 50 µg/mL streptomycin and 50 ng/mL recombinant macrophage colony stimulating factor (MCSF; PeproTech) for 11 to 14 days. Experiments were approved by the Ethics Committee of the Christian-Albrechts-University of Kiel (Kiel, Germany), in accordance with the Declaration of Helsinki.