Cells were trypsinized, counted, resuspended in medium containing 0.5% serum and then transferred to 8-μm cell culture inserts (BD Falcon) placed into 24-well plates containing complete medium. After 16 h, cells were removed from the top of the filter using a cotton swab, and migrating cells on the bottom of the filter were fixed with 4% PFA and stained with 5% crystal violet26 (link).
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