Cell Migration Assay

Materials to be prepared beforehand

1. FBS free medium
2. 10% FBS medium
3. Cell migration filter insert ( Transwell^®, 12mm Diameter, 12 μm Pore Size.)

Procedures

1. Coat the underside of filter inserts with 20μg/ml of fibronectin overnight at 4 ^oC Air dry the insert before the cells were added.
2. Trypsinize cells and wash twice with FBS free medium.
3. Count cells to 4×10⁵/ml.
4. Add 0.5ml cells in FBS free medium 10 the top of insert .
5. Add 1ml of medium (with 10% FBS and 10μg/ml Collagen I ) to the lower chamber .
6. Put filter insert into the lower chamber and incubate overnight at 37 ^oC.
7. Remove cells on the topside of the filter by scrubbing twice with cotton tipped swab moistened with no FBS medium.
8. Stain cells with Diff-Quik^TM stain kit.
9. Wash with distilled water.
10. Mount.