In Vitro Wound Healing Assays to Investigate Epidermal Migration

Re-epithelialization after cutaneous injury is a complex and multifaceted process that incorporates numerous cellular components interacting in a myriad of pathways. One of the most crucial aspects of this process is the initiation of keratinocyte migration to fill the wound bed. Re-epithelialization involves both the individual and collective movement of epidermal cells under the control of integrated signaling paradigms. It is therefore essential to develop a simple methodology to dissect the basic movement of epidermal cells in vitro. Scratch assays are relatively simple experiments in which a single layer of cells are plated onto a prepared dish with multiple furrows created in the cell bed. The resulting cellular migration to fill the wound bed can then be imaged and processed quantitatively to investigate migration rates and other factors of interest. Here, we provide important adaptations to the classic scratch assay to make it a robust, reproducible, and quantitative tool for the evaluation of epidermal cell migration.