Summary
We describe an in vivo invasion assay that enables the collection of invasive cells from the primary tumor. In addition to determination of the endogenous, unstimulated invasive properties of cells in vivo, the assay can take advantage of the chemotactic properties of cancer cells. Microneedles are filled with a mixture of extracellular matrix components such as Matrigel with or without a chemoattractant such as EGF, and then introduced into the primary tumor of a rat or mouse that is generated either by orthotopic injection of carcinoma cell lines or by a transgene such as polyoma Middle T. Over the course of 4 h the invasive cell population enters the needles while the animal is kept under anesthesia. At the end of the collection time, the invasive cells are extruded from the microneedles and can be analyzed in terms of the number and type of cells that invade in response to defined stimuli. By including pharmacological inhibitors in the needle, signaling pathways contributing to in vivo invasion can also be identified. This assay leads to a better understanding of the cell types and signaling involved in the tumor microenvironment, and has the potential to be applied to a variety of in vivo models.
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Acknowledgments
This work was supported by NIH CA110269 (LH), NIH CA100324 (JC, JES, JW), and NIH CA77522 (TS and JES). J.E.S. is the Betty and Sheldon Feinberg Senior Faculty Scholar in Cancer Research. Authors LH and TS contributed equally.
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© 2009 Humana Press
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Hernandez, L., Smirnova, T., Wyckoff, J., Condeelis, J., Segall, J.E. (2009). In Vivo Assay for Tumor Cell Invasion. In: Jin, T., Hereld, D. (eds) Chemotaxis. Methods in Molecular Biology™, vol 571. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-198-1_15
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DOI: https://doi.org/10.1007/978-1-60761-198-1_15
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