Biocoat gfr matrigel invasion chambers

For cell invasion assays, 3 × 10⁴ to 2 × 10⁵ cells were plated in an 8-μm 24-well plate chamber insert (Corning BioCoat GFR Matrigel Invasion Chambers, 354483) with serum-free medium on the top of the insert and medium containing 10% FBS in the well. Each sample was performed in triplicate. Cells were incubated for 24 hours and fixed with 4% paraformaldehyde for 15 min. After washing with PBS, cells on the top of the insert were scraped with a cotton swab. Cells that adhered to the well were stained with 0.5% crystal violet blue for 15 min and examined under the microscope. Cell migration was investigated by cell wound-healing assays. Briefly, a line was scratched through confluent cultures in six-well plates. The gap was inspected by microscopy over time as the cells moved in and filled the damaged area.

For cell migration, 2×10⁴ to 2×10⁵ cells were plated in an 8.0 µm 24-well plate chamber insert (Corning, 3422) with serum-free medium on the top of insert and 3T3 conditional medium containing 10% FBS at the bottom of the insert. Cells were incubated for 24 hr and fixed with 4% paraformaldehyde for 15 min. After washed with PBS, cells on the top of the insert were scraped with a cotton swab. Cells adherent to the bottom were stained with 0.5% crystal violet blue for 15 min and then washed with ddH₂O. The positive staining cells were examined under microscope. For cell invasion assay, Corning BioCoat GFR Matrigel Invasion Chambers (354483) were used instead of chamber insert used in migration assay. The following steps were performed as migration assay described above.