24 well transwell chambers plates

Cell migration assays were performed using transwell. Cells (1 × 10⁵) in culture medium containing 1% FBS were added to the top chamber of 24-well transwell chambers plates (8.0 μm, Corning, NY, USA.) and culture medium with 10% FBS was added in the lower chamber as a chemoattractant.
After incubation for 24 h, the underside cells of the membrane were fixed, stained with 0.1% Crystal Violet, and then the percentages of migrated cells were counted (10 random 200 × fields per well). Three independent experiments were performed, and the data were presented as the mean ± SEM. For wound healing assays, a total of 1 × 10⁶ cells/well were plated into 6-well plates and cultured to about 80% confluence. After serum starvation for 24 h, scratches were performed in the middle slides using 10 μl pipette tip for each well. The closure of the gap distance was observed and photographed at 0 h, 12 h, 24 h, 36 h and 48 h under an inverted light microscope and quantitatively evaluated using ImageJ. Each experiment was repeated at least three times.

Cells were starved in serum-free media for 24 h then added to the top chamber of 24-well transwell chambers plates (8.0 μm, Corning, NY, USA.). Specially, for the invasion assay, cells were seeded into the top chamber coated with Matrigel (BD Biosciences). Complete medium was added to the bottom chambers to stimulate migration or invasion. After incubation for 24–48 h, the cells those adhered to the lower surface of the membrane were stained with 0.1% Crystal Violet, and then the percentages of migrated or invasion cells were calculated. Five randomly selected fields per filter were counted.
In wound healing assay, cells were seeded at a density of 1 × 10⁶ cell/well onto six-well plates and cultured to about 80% confluence. Then, a sterile 10-μl pipette tip was used to form artificial scratches for each well. The suspended cells were washed away with PBS, and then the cells were cultured in medium with 1% FBS. Cell migration distance was photographed at 0 h and 24 h under an inverted light microscope.

Cells were starved in serum-free media for 24 h then added to the top chamber of 24-well transwell chambers plates (8.0 μm, Corning, NY, USA.). Specially, for the invasion assay, cells were seeded into the top chamber coated with Matrigel (BD Biosciences). Complete medium was added to the bottom chambers to stimulate migration or invasion. After incubation for 24–48 h, the cells those adhered to the lower surface of the membrane were stained with 0.1% Crystal Violet, and then the percentages of migrated or invasion cells were calculated. Five randomly selected fields per filter were counted.