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Moving cell fronts are an essential feature of wound healing, development and disease. The rate at which a cell front moves is driven, in part, by the cell motility, quantified in terms of the cell diffusivity  D , and the cell proliferation rate  λ . Scratch assays are a commonly reported procedure used to investigate the motion of cell fronts where an initial cell monolayer is scratched, and the motion of the front is monitored over a short period of time, often less than 24 h. The simplest way of quantifying a scratch assay is to monitor the progression of the leading edge. Use of leading edge data is very convenient because, unlike other methods, it is non-destructive and does not require labelling, tracking or counting individual cells among the population. In this work, we study short-time leading edge data in a scratch assay using a discrete mathematical model and automated image analysis with the aim of investigating whether such data allow us to reliably identify  D  and  λ . Using a naive calibration approach where we simply scan the relevant region of the ( D ,  λ ) parameter space, we show that there are many choices of  D  and  λ  for which our model produces indistinguishable short-time leading edge data. Therefore, without due care, it is impossible to estimate  D  and  λ  from this kind of data. To address this, we present a modified approach accounting for the fact that cell motility occurs over a much shorter time scale than proliferation. Using this information, we divide the duration of the experiment into two periods, and we estimate  D  using data from the first period, whereas we estimate  λ  using data from the second period. We confirm the accuracy of our approach using  in silico  data and a new set of  in vitro  data, which shows that our method recovers estimates of  D  and  λ  that are consistent with previously reported values except that that our approach is fast, inexpensive, non-destructive and avoids the need for cell labelling and cell counting.

How much information can be obtained from tracking the position of the leading edge in a scratch assay?