Matrigel invasion chamber 24 well plates with 8.0 μm pores

Cells were serum-starved for 24 h, then 5 × 10⁴ cells were seeded in the upper chamber (8 μm pore size, Transwell Chambers with a track-etched membrane (Corning, Inc).and Matrigel™ Invasion Chamber (BD Bioscience)) with serum-free medium, while complete medium was in the bottom chamber. Forty-eight hours later, the cells were migrated into the lower chamber and stained with 0.1% crystal violet. Migrated cells from five fields in each sample were counted for quantitative analysis. by using 24-well, New York, CA, USA).
A cell invasion assay was also conducted in the same manner but with Matrigel™ Invasion Chamber 24-well plates with 8.0-μm pores (BD Biosciences, San Jose, USA) and an incubation time of 48 h.

Cell migration was determined by using 24-well Transwell Chambers with an 8-μm pore size and a track-etched membrane (Corning, Inc., New York, CA, USA). A chamber insert was placed into each well of a 24-well dish containing 600 μl of DMEM supplemented with 20% FBS in the lower chamber. Approximately 5×10⁴ cells were suspended in 200 μl of serum-free DMEM and seeded into the upper chamber. The cells were incubated at 37 °C with 5% CO₂ for 24 h. Then, the cells on the lower chamber membrane were fixed with 100% methanol for 30 min at room temperature and stained with 0.1% crystal violet for 20 min at room temperature. Subsequently, the nonmigrated cells on the upper side of the membranes were removed, and the migrated cells on the underside of the membranes were observed with a TS100/100-F inverted fluorescence microscope (Nikon, Corporation, Tokyo, Japan) using five randomized fields. A cell invasion assay was also conducted in the same manner but with Matrigel™ Invasion Chamber 24-well plates with 8.0-μm pores (BD Biosciences, San Jose, USA) and an incubation time of 48 h.