Cell Migration Assay
Materials to be prepared beforehand
- FBS free medium
- 10% FBS medium
- Cell migration filter insert ( Transwell®, 12mm Diameter, 12 μm Pore Size.)
- Coat the underside of filter inserts with 20μg/ml of fibronectin overnight at 4 oC Air dry the insert before the cells were added.
- Trypsinize cells and wash twice with FBS free medium.
- Count cells to 4×105/ml.
- Add 0.5ml cells in FBS free medium 10 the top of insert .
- Add 1ml of medium (with 10% FBS and 10μg/ml Collagen I ) to the lower chamber .
- Put filter insert into the lower chamber and incubate overnight at 37 oC.
- Remove cells on the topside of the filter by scrubbing twice with cotton tipped swab moistened with no FBS medium.
- Stain cells with Diff-QuikTM stain kit.
- Wash with distilled water.