Abstract
Cell migration, essential in cancer progression, is a complex process comprising a number of spatiotemporally regulated and well-coordinated mechanisms. In order to study (random) cell migration in the context of responses to various external cues (such as growth factors) or intrinsic cell signaling, a number of different tools and approaches have been developed. In order to unravel the key pathways and players involved in the regulation of (cancer) cell migration, a systematical mapping of the players/pathways is required. For this purpose, we developed a cell migration assay based on automatic high-throughput microscopy screen. This approach allows for screening of hundreds of genes, e.g., those encoding various kinases and phosphatases but can also be used for screening of drugs libraries. Moreover, we have developed an automatic analysis pipeline comprising of (a) automatic data acquisition (movie) and (b) automatic analysis of the acquired movies of the migrating cells. Here, we describe various facets of this approach. Since cell migration is essential in progression of cancer metastasis, we describe two examples of experiments performed on highly motile (metastatic) cancer cells.
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Acknowledgments
The MTLn3-ErbB1 cells are a kind gift from Jeffrey Segall and the H1299-GFP from Benjamin Geiger. This work is financially supported by the Dutch Cancer Society (UL 2007–3860) and the EU FP7 Health Program Metafight (EU201862).
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van Roosmalen, W., Le Dévédec, S.E., Zovko, S., de Bont, H., van de Water, B. (2011). Functional Screening with a Live Cell Imaging-Based Random Cell Migration Assay. In: Wells, C., Parsons, M. (eds) Cell Migration. Methods in Molecular Biology, vol 769. Humana Press. https://doi.org/10.1007/978-1-61779-207-6_29
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DOI: https://doi.org/10.1007/978-1-61779-207-6_29
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