Falcon 8.0 μm pore 24 transwell cell culture inserts

For migration assay, 0.65 ml of complete culture medium was added into the lower chamber as a chemoattractant and suspended 1 × 10⁴ cells (0.2 ml) with serum free medium were layered in the upper compartment of the BD Falcon 8.0-μm pore 24-transwell cell culture inserts (BD Biosciences, Franklin lakes, NJ, USA) with duplicate. After incubation for 24 h at 37 °C in a humidified incubator, non-migrating cells were removed from the upper side of the membrane by wiping with a cotton swab. Cells at the lower side of membrane were fixed with 100% cold-methanol and stained with 0.05% crystal violet (Sigma-Aldrich, St. Louis, MO, USA). The pictures of transwell insert membrane were captured at microscope under 12.5× magnification and the total number of migrated cells were counted by AlphaEaseFC software. For invasion assay, the BD Falcon 8.0-μm pore 24-transwell cell culture inserts coated with 100 μl of 0.3 mg/ml Matrigel (BD Biosciences, Bedford, MA, USA) were re-hydrated for 2 h at 37 °C. 0.2 ml of 2 × 10⁴ cells with serum free medium were layered in the upper compartment of the transwell inserts and then the assays were performed as described in migration assay.