Wound-healing and cell migration assay

For the wound-healing experiment, cells were transplanted and grown until they developed a monolayer of 90% fusion. Cells were then removed with the tip of an aseptic straw and treated in FBS-free medium according to the instructions. The scratched visual field was randomly selected under the microscope, and the migration distance of the cells to the scratched area was measured.

Transwell system was utilized for migration analysis. And DMEM containing 10% FBS was served as a chemical inducer. Cells were fixed with 4% paraformaldehyde at 4 °C for 30 min after 18 h of incubation at 37 °C and 5% CO₂. Cells were stained with crystal violet after rinsing with PBS and incubated for 45 min at room temperature. The cells on the cell culture insert’s upper side were erased with a cotton swab, and washed thoroughly with PBS. The average number of cells was calculated according to the random visual field. Each sample was counted for three chambers.