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.