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

Abstract

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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Figure 1: Analysis of primary EC migration by in vitro scratch assay.
Figure 2: Measurement of individual cell migration in in vitro scratch assay.

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References

  1. Todaro, G.j. et al. The initiation of cell division in a contact-inhibited mammalian cell line. J. Cell Physiol. 66, 325–333 (1965).

    Article  CAS  PubMed  Google Scholar 

  2. Haudenschild, C.C. et al. Endothelial regeneration. II. Restitution of endothelial continuity. Lab. Invest. 41, 407–418 (1979).

    CAS  PubMed  Google Scholar 

  3. Etienne-Manneville, S. et al. Integrin-mediated activation of Cdc42 controls cell polarity in migrating astrocytes through PKCzeta. Cell 106, 489–498 (2001).

    Article  CAS  PubMed  Google Scholar 

  4. Fukata, Y. et al. Phosphorylation of adducin by Rho-kinase plays a crucial role in cell motility. J. Cell Biol. 145, 347–361 (1999).

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. Abbi, S. et al. Regulation of focal adhesion kinase by a novel protein inhibitor FIP200. Mol. Biol. Cell. 13, 3178–3191 (2002).

    Article  CAS  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by NIH grants GM48050 and HL73394 to J.-L. Guan.

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Correspondence to Jun-Lin Guan.

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The authors declare no competing financial interests.

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Liang, CC., Park, A. & Guan, JL. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2, 329–333 (2007). https://doi.org/10.1038/nprot.2007.30

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