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Nat Protoc. 2007;2(2):329-33.

In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro.

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Division of Molecular Medicine and Genetics and Cell and Developmental Biology, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.


The in vitro scratch assay is an easy, low-cost and well-developed method to measure cell migration in vitro. The basic steps involve creating a "scratch" in a cell monolayer, capturing the images at the beginning and at regular intervals during cell migration to close the scratch, and comparing the images to quantify the migration rate of the cells. Compared to other methods, the in vitro scratch assay is particularly suitable for studies on the effects of cell-matrix and cell-cell interactions on cell migration, mimic cell migration during wound healing in vivo and are compatible with imaging of live cells during migration to monitor intracellular events if desired. Besides monitoring migration of homogenous cell populations, this method has also been adopted to measure migration of individual cells in the leading edge of the scratch. Not taking into account the time for transfection of cells, in vitro scratch assay per se usually takes from several hours to overnight.

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