Abstract
Cells within a tumor interact by generating, transmitting, and sensing mechanical forces. Among all the cells of the tumor microenvironment, cancer-associated fibroblasts (CAFs) are a paradigmatic example of mechanical communication. In different steps of tumor progression, CAFs pull and push on cancer cells, regulating cancer cell migration, invasion, compartmentalization, and signaling. There is thus an increasing need to experimentally address mechanical interactions within a tumor. A common technique to measure these interactions is laser ablation. Cutting a tissue region with a high-power laser triggers a sudden tissue displacement whose direction and magnitude reveal the local mechanical stresses. In this chapter, we provide a detailed protocol to perform laser ablations in vitro and ex vivo. First, we describe how to prepare cocultures of primary CAFs and cancer cells and tumor explants. Then, we explain how to perform laser ablations in these two systems and how to analyze the induced tissue displacements using particle image velocimetry (PIV). Overall, we provide a workflow to perform, analyze, and interpret laser ablations to explore tumor mechanical interactions.
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Acknowledgments
We thank the Cell and Tissue Imaging Facility (PICT-IBiSA) at Institut Curie, and specially O. Renaud for technical assistance, as well as Matic-Vignjevic and Trepat labs for sharing continuous discussions, expertise, and scripts. This work was supported by the Juan de la Cierva grant program to Jorge Barbazan (IJC2018-036875-I, Ministerio de Ciencia e Innovación), and Fondation ARC pour la Recherce sur le Cancer to Carlos Perez Gonzalez.
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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Perez-Gonzalez, C., Barbazán, J. (2024). Addressing the Mechanical Interaction Between Cancer-Associated Fibroblasts and Cancer Cells by Laser Ablation. In: Sumbalova Koledova, Z. (eds) 3D Cell Culture. Methods in Molecular Biology, vol 2764. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3674-9_13
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DOI: https://doi.org/10.1007/978-1-0716-3674-9_13
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Online ISBN: 978-1-0716-3674-9
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