Cell migration
Monocyte migration testing was performed with 5μm pore-size
polycarbonate membrane transwell tissue culture inserts (Corning, USA).
Cells (5 × 104 per well) were seeded into the wells in
the top chamber in 10% FBS medium, and CXCL2 was added to the lower
chamber in 10% FBS medium. After incubation for 1 hour at 37°C the
medium was removed and the filters were fixed with 4% paraformaldehyde
for 20 minutes. The filters were then stained with 0.1% crystal violet
for 15 minutes. The number of cells that had migrated from the upper
surface of the filter to the lower surface of the filter was counted.