Human umbilical vein ecs

Human umbilical vein ECs were purchased from Lonza Incorporated (Allendale, NY, USA) and were cultured on gelatin-coated plates in M199 medium supplemented with 20% bovine calf, 2 mM L-glutamine, 50 µg/mL EC growth factor, 90 µg/mL heparin and 100 U/mL penicillin and streptomycin (GLS1 paper). The human monocytic cell line U937 (American Type Culture Collection, Manassas, VA, USA) was grown in suspension in RPMI-1640 medium containing 2 mM L-glutamine, 1 mM sodium pyruvate, 4.5 g/L glucose, 10% fetal bovine serum, and 100 µM penicillin and streptomycin. All cells were grown in an atmosphere of 95% air and 5% CO₂ at 37 °C [74 (link)].

GFP-A549 (Sigma) cells were cultured in F-12k medium (Thermo Fisher Scientific) supplemented with 10% FBS and 1% penicillin-streptomycin solution and used for no more than 15 passages. SK-MES-1 (Cell Lines Service, CLS) cells were cultured in ATCC-formulated Eagle's Minimum Essential Medium (ATCC), supplemented with 10% FBS and 1% penicillin-streptomycin solution and used for no more than 10 passages. Human Umbilical Vein ECs (Lonza) were cultured in Endothelial Growth Medium (EGM-2MV, Lonza), and passages 6 to 8 were used for the experiments. Normal Human Lung Fibroblasts (NFs, Lonza) were cultured in Fibroblast Growth Medium (FGM-2, Lonza) and used up to passage 8 for all the experiments. Human Lung CAFs were cultured in DMEM, supplemented with 10% FBS and 1% penicillin-streptomycin solution and used up to 15 passages. All cells were maintained at 37 ᵒC and 5% CO₂ in humid incubators.