Gelatin type a

Human umbilical vein ECs (HUVECs) were obtained from collagenase-digested umbilical cord veins[20 (link)]. Human aortic endothelial cells (HAECs) were isolated from aortic explants of heart transplant donors of anonymous origin through the UCLA transplant program as previously described [14 , 21 (link)]. No individual information, including ethnicity, history, and disease status of donors is known[22 (link)]. HUVECs and HAECs were cultured in Petri dishes or flasks coated with 0.2% gelatin type A (#901771; MP Biomedicals, Santa Ana, CA), in Endothelial Cell Medium (ECM, #1001, ScienCell, Carlsbad, CA) containing 465 mL of basal medium, 25 mL of FBS (#0025, ScienCell), 5 mL of Endothelial Cell Growth Supplement (ECGS, #1052, ScienCell) and 5 mL of penicillin/streptomycin solution (P/S, #0503, ScienCell). HUVECs were used in experiments between 3 and 7 passages as described previously[14 ]. HAECs were less than 15 passages. Mouse lung endothelial cells (MLECs) and mouse aortic endothelial cells (MAoECs) were isolated as described previously[19 (link)]. Cells were characterized by anti-CD31 and used for experiments within 2–3 passages as previously described[14 ].