24 well transwell chamber inserts

Cell migration and invasion potencies were studied by wound-healing and Transwell. In wound-healing assay, gastric cancer cells were seeded into 6-well plates and grow reach to 100% confluence, the single layer cells were scratched straightly with pipette tip. At 0 hour and 24 hours the distance covered by migrated cells was identified. 24-well Transwell chamber inserts (Corning, Corning, NY, USA) and Corning BioCoat Growth Factor Reduced Matrigel Invasion Chambers (Corning) were used respectively in migration and invasion assays. 6 × 10⁴ HGC 27 and 4 × 10⁴ MGC 803 were resuspended in 200 μL RPMI 1640 medium and laid into upper chamber, meanwhile 750 μL RPMI 1640 with 10% FBS was added into lower chamber. The plate was incubated for 24 hours at 37 °C in humidified incubator containing 5% CO₂. Afterwards, medium was discarded, cells were fixed with paraformaldehyde for 15 minutes and then stained with crystal violet for 15 minutes. Random fields were captured under microscope at 100×.

Transwell migration and invasion assay were performed in SGC-7901 and MGC-803 cells using 24-well transwell chamber inserts (Corning Inc., Corning, NY, USA). Briefly, cells were resuspended in serum-free medium and seeded in the upper chambers with or without Matrigel (BD Biosciences, San Jose, CA, USA). The lower chamber was filled with medium containing 10% FBS as a chemoattractant. After 24 h incubation, the cells that migrated or invaded into the lower chamber were fixed with 70% ethanol for 30 min and stained with 0.1% crystal violet for 10 min, followed by quantification in randomly selected five fields under a microscope.