Abstract
In many solid tumors, collective cell invasion prevails over single-cell dissemination strategies. Collective modes of invasion often display specific front/rear cellular organization, where invasive leader cells arise from cancer cell populations or the tumor stroma. Collective invasion involves coordinated cellular movements which require tight mechanical crosstalk through specific combinations of cell–cell interactions and cell–matrix adhesions. Cancer Associated Fibroblasts (CAFs) have been recently reported to drive the dissemination of epithelial cancer cells through ECM remodeling and direct intercellular contact. However, the cooperation between tumor and stromal cells remains poorly understood. Here we present a simple spheroid invasion assay to assess the role of CAFs in the collective migration of epithelial tumor cells. This method enables the characterization of 3D spheroid invasion patterns through live cell fluorescent labeling combined with spinning disc microscopy. When embedded in extracellular matrix, the invasive strands of spheroids can be tracked and leader/follower organization of CAFs and cancer cells can be quantified.
Sefora Conti and Takuya Kato are co-authors.
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Acknowledgements
This study was supported by MINECO/FEDER (PGC2018-099645-B-I00 to X.T.), the Generalitat de Catalunya (SGR-2017-01602 to X.T. and CERCA Program), the European Research Council (CoG-616480 to X.T.), the European Commission (Grant Agreement SEP-210342844 to X.T.). A.L. has received financial support through the Junior Leader Postdoctoral Fellowship Programme from “la Caixa” Banking Foundation (LCF/BQ/PR18/11640001). T.K. is funded by Marie-Curie action (HeteroCancerInvasion # 708651) and the Japanese Strategic Young Researcher Overseas Visits Program for Accelerating Brain Circulation. D.P is supported by Breast Cancer Now (BCN) (2013NovPR182/Breast Cancer Now (BCN)). S.C is supported by the 2017 FPI MINECO doctoral fellowship programme (BES-2017-079847). E.S. is supported by the Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001144), the UK Medical Research Council (FC001144) and the Wellcome Trust (FC001144). The authors wish to acknowledge the role of the Breast Cancer Now Tissue Bank in collecting and making available the samples used in the generation of this publication. IBEC is recipient of a Severo Ochoa Award of Excellence from the MINECO.
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Conti, S., Kato, T., Park, D., Sahai, E., Trepat, X., Labernadie, A. (2021). CAFs and Cancer Cells Co-Migration in 3D Spheroid Invasion Assay. In: Campbell, K., Theveneau, E. (eds) The Epithelial-to Mesenchymal Transition. Methods in Molecular Biology, vol 2179. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0779-4_19
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DOI: https://doi.org/10.1007/978-1-0716-0779-4_19
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