A Robust and Standardized Approach to Quantify Wound Closure Using the Scratch Assay

The scratch assay is an in vitro assay that allows for high-throughput quantification of wound closure by keratinocytes and fibroblasts with relative ease. However, this assay is amenable to experimental variables, which can result in false-positive and false-negative data, making the interpretation...

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Bibliographic Details
Published in:Methods and protocols 2023-09, Vol.6 (5), p.87
Main Authors: Balko, Stefan, Kerr, Evan, Buchel, Edward, Logsetty, Sarvesh, Raouf, Afshin
Format: Article
Language:English
Subjects:
Cell cycle
dermal fibroblasts
Fibroblasts
Growth factors
Keratinocytes
Protocol
Wound healing
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Summary:The scratch assay is an in vitro assay that allows for high-throughput quantification of wound closure by keratinocytes and fibroblasts with relative ease. However, this assay is amenable to experimental variables, which can result in false-positive and false-negative data, making the interpretation of such data difficult. Also, data variability decreases the sensitivity of the scratch assay. Here, we identify important sources of data variation in the scratch assay and provide rational mitigation strategies that enable robust and highly reproducible quantification of scratch width and area, and ultimately the scratch closure rates. By eliminating these sources of variability, the sensitivity of the scratch assay is enhanced, thereby allowing for identification of dependent variables with wide-ranging impacts on wound closure in a robust and standardized manner.
ISSN:2409-9279
2409-9279
DOI:10.3390/mps6050087