Cell Invasion and Migration Assay

The cell invasion assay was performed using 24-well transwell insert chambers with a polycarbonate filter with a pore size of 8.0 µm (Costar, Corning, NY). A total of 2×10⁴ cells in serum-free medium were seeded on top of a Matrigel-coated filter (BD Biosciences, Franklin Lakes, NJ) in each insert chamber. Then, the insert chambers were placed into wells on top of culture medium containing 10% horse serum as a chemoattractant. The migration assay was performed using 24-well transwell insert chambers with a polycarbonate filter without Matrigel. The invasive or migratory ability of cells was determined by the number of cells translocated to the lower side of filters [35] (link), [36] (link).