24 well matrigel coated transwell chambers

The cell invasion ability was examined using 24-well matrigel-coated Transwell chambers (BD Biosciences) as previously described (6 (link)). In brief, the SW1116 and HCT116 cells were treated with Form for 48 h, washed with PBS and resuspended at 1×10⁵ cells/ml in serum-free medium. Then 0.2 ml cell suspension was added to the upper chamber and 0.5 ml medium containing 10% FBS was added to the bottom chamber. Following 24 h incubation, all non-invaded cells were removed from the upper face of the filters and the invaded cells were fixed with 4.0% paraformaldehyde at room temperature for 15 min and stained by 0.1% crystal violet solution at room temperature for 2 min. The experiments were repeated in triplicate wells and the invaded cells were counted using a light microscope (×400) in five different fields of view per filter.

The invasion ability of JEG-3 cells was evaluated using the 24-well Matrigel-coated
Transwell chambers (BD Biosciences). The upper chamber was filled with 1 × 10⁵cells/mL in FBS-free DMEM, and the lower chamber was filled with 600 μL of DMEM containing
10% FBS. After incubation at 37°C for 24 hours, cells on the upper chambers were removed,
and invaded cells on the lower surface of the chambers were fixed in 100% methanol and
stained with 0.1% crystal violet. Invaded cell was counted and photographed under a
microscope (×200 magnification, Nikon TE2000).