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.