Cell Migration Assay Using Boyden Chamber

Cell migration was analyzed by a Boyden chamber assay utilizing ThinCert® transwell polyethylene terephthalate (PET) membrane supports (8 µm pore size; Greiner Bio-One, St. Gallen, Switzerland) as described (Asparuhova et al., 2021 (link)). After 24 h of starvation, 3 × 10⁴ cells were cultured in the top insert chamber with 200 µl 0% FCS/DMEM. Each of the collagen matrices was placed in the low chamber with 800 µl 10% FCS/DMEM. Cells were allowed to migrate across the capillary pore PET membrane for 18 h at 37°C before fixation in Shandon™ Formal-Fixx™ (ThermoFisher Scientific), and staining in 0.1% crystal violet solution (Sigma). Images of duplicate inserts were acquired on an Olympus CKX41 microscope using a ProgResCT3 camera. Migration was quantified by using the ImageJ software (Schneider et al., 2012 (link)) as described (Gurbuz et al., 2014 (link)). Data represent means ± SD from three independent experiments performed with each of the two cell lines.