Macrophage Differentiation and Isolation

Bone marrow cells were differentiated into mature macrophages (BMDMs) from wild type C57BL/6 (WT) and NFATc3^−/− mice in DMEM supplemented with 10% FBS, 1% penicillin/streptomycin and 20 ng/mL recombinant mouse M-CSF added on Day 1, 3 and 5. Cells were differentiated up to 7 days in total [4 (link)–6 (link)]. Mouse lung total macrophages were isolated by collagenase digestion, purified by adherence for 1 hour, plated in DMEM supplemented with 10% FBS, 1% penicillin/streptomycin [51 (link)]. Mouse PMVEC were isolated by Miltenyi CD31 magnetic beads. AEC type II cells were isolated as described earlier [52 ].