<i>In vitro</i> cell migration quantification method for scratch assays

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  • Ana Victoria Ponce Bobadilla
    Institute for Applied Mathematics, Heidelberg University, 69120 Heidelberg, Germany
  • Jazmine Arévalo
    Renal Physiopathology Group, CIBBIM-Nanomedicine, Vall d’Hebron Research Institute, Barcelona, Spain
  • Eduard Sarró
    Renal Physiopathology Group, CIBBIM-Nanomedicine, Vall d’Hebron Research Institute, Barcelona, Spain
  • Helen M. Byrne
    Wolfson Centre for Mathematical Biology, Mathematical Institute, University of Oxford, Oxford OX2 6GG, UK
  • Philip K. Maini
    Wolfson Centre for Mathematical Biology, Mathematical Institute, University of Oxford, Oxford OX2 6GG, UK
  • Thomas Carraro
    Institute for Applied Mathematics, Heidelberg University, 69120 Heidelberg, Germany
  • Simone Balocco
    Department of Mathematics and Informatics, University of Barcelona, Gran Via 585, 08007 Barcelona, Spain
  • Anna Meseguer
    Renal Physiopathology Group, CIBBIM-Nanomedicine, Vall d’Hebron Research Institute, Barcelona, Spain
  • Tomás Alarcón
    ICREA, Pg. Lluís Companys 23, 08010 Barcelona, Spain

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Description

<jats:p> The scratch assay is an <jats:italic>in vitro</jats:italic> technique used to assess the contribution of molecular and cellular mechanisms to cell migration. The assay can also be used to evaluate therapeutic compounds before clinical use. Current quantification methods of scratch assays deal poorly with irregular cell-free areas and crooked leading edges which are features typically present in the experimental data. We introduce a new migration quantification method, called ‘monolayer edge velocimetry’, that permits analysis of low-quality experimental data and better statistical classification of migration rates than standard quantification methods. The new method relies on quantifying the horizontal component of the cell monolayer velocity across the leading edge. By performing a classification test on <jats:italic>in silico</jats:italic> data, we show that the method exhibits significantly lower statistical errors than standard methods. When applied to <jats:italic>in vitro</jats:italic> data, our method outperforms standard methods by detecting differences in the migration rates between different cell groups that the other methods could not detect. Application of this new method will enable quantification of migration rates from <jats:italic>in vitro</jats:italic> scratch assay data that cannot be analysed using existing methods. </jats:p>

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