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Transwell cell migration and invasion assay

YL Yu-Mei Liao
YW Ya-Hui Wang
JH Jung-Tung Hung
YL Yu-Ju Lin
YH Yen-Lin Huang
GL Guo-Shiou Liao
YH Ya-Ling Hsu
JW Jen-Chien Wu
AY Alice L. Yu
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For migration assay, 4 × 104 cells in 100 μl serum-free medium were seeded on top of the filter membrane (8.0 μm pores) in a transwell insert (6.5 mm diameter) (Corning, NY, USA). 10% FBS (v/v) was supplemented in medium and placed in the lower chamber as a chemoattractant. Cells were allowed to migrate at 37 °C for 15 h, and the transwell insert was removed from the plate for fixation and staining with crystal violet. The migrated cells on the underside of the filter membrane were viewed with an inverted microscope and counted for cell numbers. For invasion assay, similar procedures were performed but with 60 μl matrigel placed onto the upper chamber of transwell and incubated at 37 °C for 1 h before seeding the cells.

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