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Studying tumor growth in Drosophila using the tissue allograft method

Abstract

This protocol describes a method to allograft Drosophila larval tissue into adult fly hosts that can be used to assay the tumorigenic potential of mutant tissues. The tissue of interest is dissected, loaded into a fine glass needle and implanted into a host. Upon implantation, nontransformed tissues do not overgrow beyond their normal size, but malignant tumors grow without limit, are invasive and kill the host. By using this method, Drosophila malignant tumors can be transplanted repeatedly, for years, and therefore they can be aged beyond the short life span of flies. Because several hosts can be implanted using different pieces from a single tumor, the method also allows the tumor mass to be increased to facilitate further studies that may require large amounts of tissue (i.e., genomics, proteomics and so on). This method also provides an operational definition of hyperplastic, benign and malignant growth. The injection procedure itself requires only 1 d. Tumor development can then be monitored until the death of the implanted hosts.

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Figure 1: Flowchart outlining the key stages of the PROCEDURE.
Figure 2: The injection equipment.
Figure 3: Dissection of larval brain and optic lobe isolation.
Figure 4: Allograft procedure.
Figure 5: Example of tumor growth and downstream application.
Figure 6: Isolation of allografted tumors.

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Acknowledgements

J. Szabad kindly hosted and demonstrated this technique to C.G. at the Department of Medical Biology, University of Szeged, in 2003. His contribution is greatly appreciated. Work in our laboratory is supported by grants AdG 2011 294603 advanced grant from the European Research Council, BFU2012–32522 from the Spanish Ministerio de Economí y Competitividad (MINECO) and Agaur 2014 SGR 100 from Generalitat de Catalunya.

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Authors

Contributions

C.G. and F.R. designed the experiments. C.G. implemented the protocol. F.R. conducted the experiments and prepared all the supporting media. C.G. and F.R. wrote the paper.

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Correspondence to Cayetano Gonzalez.

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The authors declare no competing financial interests.

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Rossi, F., Gonzalez, C. Studying tumor growth in Drosophila using the tissue allograft method. Nat Protoc 10, 1525–1534 (2015). https://doi.org/10.1038/nprot.2015.096

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