Description Various phenotypic characteristics are required for a cancer cell to successfully complete the metastatic cascade. Among these, acquisition of a motile phenotype is one requirement for a cell to become metastatically competent. Procedure 1.Place the chamber in 6 well tissue culture plate. Plate 1 million cells in the top chamber of noncoated polyethylene teraphthalate membranes (6-well insert, pore size 8 mm; Becton Dickinson). 2.Incubate cells for 24 h at 37oC. 3. Remove the cells that did not migrate through the pores in the membrane by scraping the membrane with a cotton swab. 4.Stain the cells that transversed the membrane with a Diff-Quick cell staining kit (Dade Behring, Inc., Newark, DE) and mount on the slide. 5.Count the motile cells in 10 random fields of view at 100X magnification 6.Express the cells as the average number of cells/field of view.
