Cellular culture and in vitro vascular permeability assays
Human Microvascular Endothelial Cells-Lung (HMVEC-L) were acquired from Lonza CC-2527 and maintained in EGM™-2MV BulletKit™. The effect of EVs in the endothelial barrier integrity was evaluated using Transwell 24-well cell culture inserts (TWs), as previously described [24]. HMVEC-L in vitro TW systems were incubated with 100µg/ml of AnEVs or BEVs in EBM-2 medium (EBM)+PBS or mediators of interest. The trans-endothelial-electrical-resistance (TEER) value of the monolayer was measured at different times (10 minutes, 1, 2 and 5 hours) through an EndOhm chamber (WPI) in individual cups.