Optimized Scratch Assay for In Vitro Testing of Cell Migration with an Automated Optical Camera

Journal of Visualized Experiments : JoVE
Michelle V Mouritzen, Håvard Jenssen

Abstract

Cell migration is an important process that influences many aspects of health, such as wound healing and cancer, and it is, therefore, crucial for developing methods to study the migration. The scratch assay has long been the most common in vitro method to test compounds with anti- and pro-migration properties because of its low cost and simple procedure. However, an often-reported problem of the assay is the accumulation of cells across the edge of the scratch. Furthermore, to obtain data from the assay, images of different exposures must be taken over a period of time at the exact same spot to compare the movements of the migration. Different analysis programs can be used to describe the scratch closure, but they are labor intensive, inaccurate, and forces cycles of temperature changes. In this study, we demonstrate an optimized method for testing the migration effect, e.g. with the naturally occurring proteins Human- and Bovine-Lactoferrin and their N-terminal peptide Lactoferricin on the epithelial cell line HaCaT. A crucial optimization is to wash and scratch in PBS, which eliminates the aforementioned accumulation of cells along the edge. This could be explained by the removal of cations, which have been shown to have an ...Continue Reading

Citations

Jun 26, 2018·Journal of Peptide Science : an Official Publication of the European Peptide Society·Michelle V MouritzenHåvard Jenssen
Sep 24, 2020·Oxidative Medicine and Cellular Longevity·Nizioł-Łukaszewska ZofiaBujak Tomasz
Jul 30, 2020·PloS One·Alejandra Suarez-ArnedoCarolina Muñoz-Camargo
Sep 19, 2019·ACS Medicinal Chemistry Letters·Angel ResendezSanjay V Malhotra

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Journal of Peptide Science : an Official Publication of the European Peptide Society
Michelle V MouritzenHåvard Jenssen
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