Wound healing migration assay

Cell migration was also analyzed by using a wound healing assay. Briefly, cells were seeded at a density of 2×10⁵ per well in a 6-well plate. At the appropriate times, cell monolayers were scratched with the tip of a standard 200-μl pipette tip. After being washed three times with PBS, cells were microphotographed (×400) and then incubated under standard conditions. Migration into the scratched area was documented at various time-points: 6, 12, 24, and 48 h after wounding. Scratch closure by migrating cells was compared in the experimental and the control cells relative to the total area of the scratches. Three to six scratches were evaluated for each experimental condition.