assessing internalized porphyromonas gingivalis in host endothelial cells

Assessing Internalized Porphyromonas gingivalis in Host Endothelial Cells

Following infection of HUVEC cells, wash the cells three times with anaerobic PBS. Next, add 2 milliliters of VEGF medium supplemented with pre-optimized concentrations of antibiotics to the HUVEC cells. Incubate the cells for one hour. Aspirate the medium, and add 2 milliliters of 1% saponin solution in BHI to each well. Incubate for 15 minutes.
Next, scrape the lysed cells, and collect the lysate in a centrifuge tube. Dilute the lysate 1 to 1 with BHI medium, and then prepare serial dilutions starting at 1 to 100. Plate 200 microliters of the suitable concentrations on blood agar plates. Allow the bacteria to grow for seven days in an anaerobic chamber, and then count the colonies.