Biocoat matrigel

Wound healing assay and transwell assay were used to assess cell migration and invasion, respectively. Briefly, for wound healing assay, cells seeded in six-well plates were cultured to 90% confluence. The monolayer cells were manually scraped using a 10-uL sterile pipette tip. The wounding scratches at 24 hr was measured and expressed as the relative percentage of the initial distance at 0 hr following formula: migration rate = migration distance/original distance. For transwell assay, cells (5×10⁴) were suspended in serum-free DMEM and added to chambers (8 mm, BD Biosciences) coated with BD BioCoat Matrigel. After incubation, the cells on the upper membrane surface were removed with a cotton tip. Then, the member was fixed and stained by violet crystalline.

Both cell migration and invasion were performed using the Transwell assay. For cell migration assay, 6  ×  10⁴ U251 cells were seeded in a serum-free medium in the upper chamber, which contained a polyethylene terephthalate membrane (with 6.5 mm in diameter and 8 μm in pore size). The bottom chamber was prepared with 10% FBS as a chemoattractant. After incubating at 37°C for 24 h, nonmigrated cells were scraped from the upper surface of the membrane with a cotton swab, and the cells migrating to the bottom chamber were fixed with paraformaldehyde and stained with crystal violet. Finally, the stained cells were counted under a microscope at  × 400 magnification in five randomly selected fields for quantification.
For cell invasion assay, 5 × 10⁴ U251 cells were suspended in 200 mL of serum-free DMEM and then treated using the same procedure as for the migration assay following the manufacturer’s protocol, but the chambers (8 mm, BD Biosciences, San Jose, CA, USA) were plated with BD BioCoat Matrigel.