2.12. Smooth Muscle Cell Wound Injury Repair Assay

After VSMCs were treated with 20 μM corylin or 5 nM sirolimus for 1 h, wounds were inflicted in their monolayers by dragging a sterile pipette tip across the monolayers. A 250-μm cell-free path was created, after which the medium was replaced with fresh medium to remove the cell debris. After being rinsed, the cells were cultured in complete medium containing the vehicle control or 20 ng/mL PDGF-BB with or without 20 μM corylin or 5 nM sirolimus. The degree of cell growth into the wound area was measured after 24 h. Time-lapse images were taken every 10 min after wounding throughout the 24-h period. The cell migration rate was measured after collection of the sequential time-lapse images. Analyses were performed on the sequential phase-contrast images with MetaMorph software (v.7.7 Molecular Devices, CA, USA).