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A transient parabiosis skin transplantation model in mice

Nature Protocols volume 7, pages 763–770 (2012)Cite this article

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Abstract

Parabiosis—conjoined surgery to provide a shared circulation between two mice—has been previously developed to study the hematopoietic system. This protocol describes the use of parabiosis for efficient transplantation of skin from a transgenic to a wild-type mouse. It can be used to study the role of stromal cells in a spontaneous model of distant cancer dissemination (metastasis). We have recently shown that primary tumor-derived stromal cells may facilitate metastasis by providing a provisional stroma at the secondary site. Studying the role of primary tumor–derived stroma cells requires methods for distinguishing and targeting stromal cells originating from the primary tumor versus their counterparts in the metastatic site. Parabiosis may also be used, taking advantage of the shared circulation between the parabiosed mice, to study tumor metastasis from one parabiont to another, or to investigate the role of circulating inflammatory cells or stem cells. Studying the role of stromal cells in metastasis using this model typically takes up to 11 weeks.

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Figure 1: Design of the experiment.
Figure 2: GFP+ skin transplantation through parabiosis surgery under general anesthesia.
Figure 3: Quantification of tumor-derived GFP+ stromal cells in lung metastases.
Figure 4: Passenger stromal cells in spontaneous metastasis.

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Acknowledgements

This work 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, S. Roberge and P. Huang with animal models.

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

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, Dan G Duda, Rakesh K Jain & Dai Fukumura

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  1. Annique M M J Duyverman
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  2. Mitsutomo Kohno
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  3. Dan G Duda
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Contributions

D.G.D., D.F. and R.K.J. designed the studies; A.M.M.J.D. 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. and D.F. wrote the manuscript.

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Correspondence to Dai Fukumura.

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The authors declare no competing financial interests.

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Duyverman, A., Kohno, M., Duda, D. et al. A transient parabiosis skin transplantation model in mice. Nat Protoc 7, 763–770 (2012). https://doi.org/10.1038/nprot.2012.032

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