Abstract
Cell migration through 3D extracellular matrices (ECMs) is crucial to the normal development of tissues and organs and in disease processes, yet adequate analytical tools to characterize 3D migration are lacking. The motility of eukaryotic cells on 2D substrates in the absence of gradients has long been described using persistent random walks (PRWs). Recent work shows that 3D migration is anisotropic and features an exponential mean cell velocity distribution, rendering the PRW model invalid. Here we present a protocol for the analysis of 3D cell motility using the anisotropic PRW model. The software, which is implemented in MATLAB, enables statistical profiling of experimentally observed 2D and 3D cell trajectories, and it extracts the persistence and speed of cells along primary and nonprimary directions and an anisotropic index of migration. Basic computer skills and experience with MATLAB software are recommended for successful use of the protocol. This protocol is highly automated and fast, taking <30 min to analyze trajectory data per biological condition.
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Acknowledgements
This work was supported in part by the US National Institutes of Health grant U54CA143868.
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A.G. conducted the experiments; P.-H.W. developed, tested, applied and validated the APRW model; and A.G., P.-H.W. and D.W. designed the experiments, analyzed the results and wrote the paper.
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The authors declare no competing financial interests.
Supplementary information
Supplementary Text and Figures
Supplementary Methods (PDF 188 kb)
Supplementary Software
Contains MATLAB scripts. (ZIP 21 kb)
Supplementary Data
Contains example excel sheets (ZIP 3455 kb)
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Wu, PH., Giri, A. & Wirtz, D. Statistical analysis of cell migration in 3D using the anisotropic persistent random walk model. Nat Protoc 10, 517–527 (2015). https://doi.org/10.1038/nprot.2015.030
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DOI: https://doi.org/10.1038/nprot.2015.030
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