Abstract
Tumor invasion is the outcome of a complex interplay between cancer cells and the stromal environment and requires the infiltration of a dense, cross-linked meshwork of collagen type I extracellular matrix. We use a membrane-free single-cell and spheroid-based complementary model to study cancer invasion through native collagen type I matrices. Cell morphology is preserved during the assays allowing real-time monitoring of invasion-induced changes in cell structure and F-actin organization. Combination of these models with computerized quantification permits the calculation of highly reproducible and operator-independent data. These assays are versatile in the use of fluorescent probes and have a flexible kinetic endpoint. Once the optimal experimental conditions are empirically determined, the collagen type I invasion assays can be used for preclinical validation of small-molecule inhibitors targeting invasion. Initiation and monitoring of the single-cell and spheroid invasion model can be achieved in 8 h (over 3 days) and in 14 h (over 5 days), respectively.
An Hendrix and Astrid De Boeck contributed equally to this work.
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Acknowledgements
M. Mareel is gratefully acknowledged for stimulating discussions. We thank Georges De Bruyne for excellent technical assistance. This work was funded by Fund for Scientific Research-Flanders (Kom op Tegen Kanker-VLK: FWOAL455) (Brussels, Belgium), INSERM, ARC, and the Scientific Exchange Program between the Flemish community and France (Grant I.2007.03; T2009.14). O. De Wever was supported by an EACR travel fellowship, and a postdoctoral grant from Fund for Scientific Research-Flanders. W. Westbroek was supported by the Intramural Research Program of the National Human Genome Research Institute.
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De Wever, O. et al. (2014). Single Cell and Spheroid Collagen Type I Invasion Assay. In: Dwek, M., Schumacher, U., Brooks, S. (eds) Metastasis Research Protocols. Methods in Molecular Biology, vol 1070. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4614-8244-4_2
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DOI: https://doi.org/10.1007/978-1-4614-8244-4_2
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