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PriCells: Transendothelial flux of primary vascular endothelial cell
PriCells: Transendothelial flux of primary vascular endothelial cell
1. Primary vascular endothelial cell (1 x 105 cells) were cultured to confluence on tissue-culture Transwell plates (polycarbonate membrane, 12-mm diameter, 0.4-µM pore size; Becton Dickinson Labware, Bedford, MA)
2. PriCells culture medium system was replaced with colorless Hanks balanced salt solution (HBSS) supplemented with 5% FBS and gentamycin (0.05 mg/ml).
3. Experimental wells were treated.
4. Control wells were treated with PBS alone.
5. Evans blue dye (20 µl of 0.5% wt/vol) was added to the upper compartment.
6. The liquid was collected from the lower wells after 30 and 60 minutes.
7. Permeability measured spectrophotometrically at 630 nm.