Abstract
Cell migration is a process that ensures correct cell localization and function in development and homeostasis. In disease such as cancer, cells acquire an upregulated migratory capacity that leads to their dissemination throughout the body. Live imaging of cell migration allows for better understanding of cell behaviors in development, adult tissue homeostasis and disease. We have optimized live imaging procedures to track cell migration in adult murine tissue explants derived from: (1) healthy gut; (2) primary intestinal carcinoma; and (3) the liver, a common metastatic site. To track epithelial cell migration in the gut, we generated an inducible fluorescent reporter mouse, enabling us to visualize and track individual cells in unperturbed gut epithelium. To image intratumoral cancer cells, we use a spontaneous intestinal cancer model based on the activation of Notch1 and deletion of p53 in the mouse intestinal epithelium, which gives rise to aggressive carcinoma. Interaction of cancer cells with a metastatic niche, the mouse liver, is addressed using a liver colonization model. In summary, we describe a method for long-term 3D imaging of tissue explants by two-photon excitation microscopy. Explant culturing and imaging can help understand dynamic behavior of cells in homeostasis and disease, and would be applicable to various tissues.
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Acknowledgments
We would like to thank Basile Gurchenkov and Fatima El Marjou for assistance with microscopes and mice. The authors greatly acknowledge the Cell and Tissue Imaging (PICT-IBiSA), Institut Curie, member of the French National Research Infrastructure France-BioImaging (ANR10-INBS-04). This work was supported by the Fondation pour la Recherche Médicale (FRM N° DGE20111123020), the Canceropole-IdF (nis2012-2-EML-04-IC-1), INCa (Cancer National Institute, n° 2011-1-LABEL-IC-4). The authors would like to acknowledge the Cell and Tissue Imaging Platform - PICT-IBiSA (member of France–Bioimaging, ANR-10-INBS-04) of the Genetics and Developmental Biology Department (UMR3215/U934) of Institut Curie for help with image analysis.
This work is funded by Institut Thématique Multi-organismes Cancer–Plan Cancer 2014–2019 and Ecole Doctorale Frontières du Vivant (FdV)–Programme Bettencourt (RS), Marie Curie Individual Fellowship (FiBRO) (JB), and ERC starting grant (DMV).
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Staneva, R., Barbazan, J., Simon, A., Vignjevic, D.M., Krndija, D. (2018). Cell Migration in Tissues: Explant Culture and Live Imaging. In: Gautreau, A. (eds) Cell Migration. Methods in Molecular Biology, vol 1749. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-7701-7_13
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DOI: https://doi.org/10.1007/978-1-4939-7701-7_13
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