Cell Invasion Assay with Matrigel

Cells were treated with 10 μg/mL mitomycin C for 2 hours and then split with trypsin/EDTA. A total of 5 × 10⁴ cells were seeded on top of 8 μm chambers without (cell culture inserts; catalog 10769-242, VWR) and with Matrigel (BioCoat Growth Factor Reduced Matrigel Invasion Chambers; catalog 354483, Corning) and then grown with 0.1% FBS-containing medium on top and 10% FBS-containing medium on the bottom (81 (link)). After 48 hours, cells were removed from the top of the filter inserts, and cells that had passed through the 8 μm pore membranes were fixed with methanol and revealed with the Harleco Hemacolor Stain Set (MilliporeSigma). Images were captured by microscopy and quantitation was performed by counting cells in 3 random fields at ×10 original magnification.