Matrigel coated transwell insert chambers

Chemotaxis of bladder cells to 10% FBS in MCoy’s medium was evaluated using a modified Boyden’s chamber with 8 μm pore polycarbonate membrane inserts (Transwell; Corning Life Sciences, PZ HTL SA, Warsaw, Poland), and the chemotactic cells were visualized with Wright’s staining (Sigma-Aldrich, Darmstadt, Germany), as previously described [64 (link)]. BSA MCoy’s medium 0.5% was used as a negative control. Before the experiment, RT4 and T24 cells were treated with sorafenib at IC50 concentration or CR-BAY aggregates for 24 h. 2.0 × 10⁴ RT4 and T24 cells in 0.5 mL in starving medium 0.5% BSA MCoy’s with or without the inhibitor were seeded per one insert. Bladder cancer cells’ invasion ability was measured in Matrigel-coated Transwell insert chambers (Corning LifeSciences, PZ HTL SA, Warsaw, Poland). The inserts were coated with 1 mg/mL Matrigel matrix according to the manufacturer’s recommendations. Methods used in the cell invasion assay were similar to those in the cell chemotactic assay.