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An isolated tumor perfusion model in mice

Nature Protocols volume 7, pages 749–755 (2012)Cite this article

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Abstract

The role of stromal cells in the tumor microenvironment has been extensively characterized. We and others have shown that stromal cells may participate in several steps of the metastatic cascade. This protocol describes an isolated tumor perfusion model that enables studies of cancer and stromal cell shedding. It could also be used to study the effects of therapies interfering with the shedding of tumor cells or fragments, circulating (stem) cells or biomarkers. Primary tumors are grown in a microenvironment in which stromal cells express GFP ubiquitously. Tumors are implanted orthotopically or can be implanted ectopically. As a result, all tumor-associated stromal cells express GFP. This technique can be used to detect and study the role of stromal cells in tumor fragments within the circulation in mice. Studying the role of stromal cells in circulating tumor fragments using this model may take 2–10 weeks, depending on the growth rate of the primary tumor.

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Figure 1: Design of the experiment.
Figure 2: Cannulation of the carotid artery.
Figure 3: Representative images of tumor cell clumps retained on a 40-mm mesh.
Figure 4: Size and viability of circulating metastatic cancer cells.

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Acknowledgements

The work of the authors is supported by US National Cancer Institute grants P01-CA80124, R01-CA115767, R01-CA85140, R01-CA126642 and T32-CA73479 (R.K.J.), R01-CA96915 (D.F.), R21-CA139168 and R01-CA159258 (D.G.D.) and Federal Share Proton Beam Program grants (R.K.J., D.F. and D.G.D.); Department of Defense Innovator Award W81XWH-10-1-0016 (R.K.J.) and Predoctoral Fellowship W81XWH-06-1-0781 (A.M.M.J.D.); American Cancer Society grant RSG-11-073-01TBG (D.G.D.); and Stichting Michael Van Vloten Fonds and the Stichting Jo Kolk (A.M.M.J.D.). We acknowledge the outstanding technical assistance of J. Kahn and P. Huang with animal models.

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Author notes

  1. Annique M M J Duyverman and Mitsutomo Kohno: These authors contributed equally to this protocol.

Authors and Affiliations

  1. Department of Radiation Oncology, Edwin L. Steele Laboratory for Tumor Biology, Massachusetts General Hospital (MGH) and Harvard Medical School, Boston, Massachusetts, USA

    Annique M M J Duyverman, Mitsutomo Kohno, Sylvie Roberge, Dai Fukumura, Dan G Duda & Rakesh K Jain

Authors
  1. Annique M M J Duyverman
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  2. Mitsutomo Kohno
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Contributions

D.G.D., D.F. and R.K.J. designed the studies; A.M.M.J.D., S.R. and M.K. performed the experiments; D.G.D., D.F., A.M.M.J.D., M.K. and R.K.J. analyzed the data; and A.M.M.J.D., D.G.D., D.F. and R.K.J. edited the manuscript.

Corresponding author

Correspondence to Rakesh K Jain.

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The authors declare no competing financial interests.

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Duyverman, A., Kohno, M., Roberge, S. et al. An isolated tumor perfusion model in mice. Nat Protoc 7, 749–755 (2012). https://doi.org/10.1038/nprot.2012.030

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