Pet membrane 24 well 8.0 μm pore size cell culture inserts

Boyden chamber migration assays were conducted using transparent PET membrane 24‐well 8.0‐μm pore size cell culture inserts (BD Falcon, Franklin Lakes, NJ, USA) coated with collagen type I‐C (Nitta Gelatin, Osaka, Japan). After the cells were seeded in triplicate on the inserts, the cells that had not invaded the lower surfaces of the filters were removed from the upper faces of the filters using cotton swabs. Cells that had invaded into the lower surfaces of the filters were fixed in methanol and acetone and stained with Giemsa. Invasion was quantitated by visually counting the photographed cells, and evaluated by statistical analysis.

Boyden chamber migration assays were conducted using transparent PET membrane 24-well 8.0 μm pore size cell culture inserts (BD Falcon, Franklin Lakes, NJ) coated with collagen type I-C (Nitta Gelatin, Osaka, Japan). After cells were seeded in triplicate on the inserts, cells that had not invaded the lower surfaces of the filters were removed from the upper faces of the filters using cotton swabs. Cells that invaded into the lower surfaces of the filters were fixed in acetone:methanol (1:1) and stained with Trypan Blue. Invasion was quantitated by visually counting photographed cells. Cell numbers were evaluated by statistical analysis.