24 well transwell chambers with 8 μm pore size polycarbonate membrane

The abilities of cell migration and invasion were tested using the 24-well transwell chambers with 8 μm pore size polycarbonate membrane (Corning, NY, USA). For cell invasion assay, the filter was pre-coated with 500 ng/ml matrigel solution (BD, Franklin Lakes, NJ, USA) and incubated at 37°C for 4 h before the invasion assay started; placed 500 μL of 10% FBS medium in the lower chamber, and placed 100 μL serum-free medium in the upper chamber. After incubation at 37°C for 24 h, the cells on the upper membrane surface were scraped off. Surface were fixed with methanol and glacial acetic acid 30 min and then stained with 10% Giemsa solution. Number of cells was counted, and five randomly fields were counted randomly in each well. The photographs were taken at 200× magnification.

For cell migration and invasion assays, 24‐well transwell chambers with 8‐μm pore size polycarbonate membrane were used (Corning). Cells were seeded on the top side of the membrane pre‐coated with Matrigel (BD) (without Matrigel for cell migration assay). Incubation for 24 hours, cells inside the upper chamber were removed with cotton swabs, while cells on the lower membrane surface were fixed and then stained with 0.5% crystal violet solution. Five fields were counted randomly in each well. All assays were performed in triplicate, and the experiment was repeated three times.