Defining Cancer Stem Cells by Xenotransplantation in Zebrafish

  • Michael C. Dovey
  • Leonard I. Zon
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 568)

Summary

The zebrafish (Danio rerio) has become an increasingly utilized and relevant model organism in the study of cancer. The use of transgenic and reverse genetic approaches has yielded several strains that model a variety of human neoplasms. In addition to modeling human disease, these strains provide a platform for the analysis of tumor stem cells. Here we describe the basic technique for the isolation and transplantation of tumor tissue in the zebrafish. This technique was designed to study metastasis and invasive potential of zebrafish tumor cells. Additionally, the basic protocol can be modified in order to describe cancer stem cell characteristics, including proliferative capacity, self-renewal, and the minimum number of tumor cells required for engraftment.

Key words

Zebrafish Cancer stem cells Transplantation Metastasis 

References

  1. 1.
    Amatruda, J.F., J.L. Shepard, H.M. Stern, and L.I. Zon, Zebrafish as a cancer model system. Cancer Cell, 2002. 1(3): p. 229–31.PubMedCrossRefGoogle Scholar
  2. 2.
    Berghmans, S., R.D. Murphey, E. Wienholds, D. Neuberg, J.L. Kutok, C.D. Fletcher, J.P. Morris, T.X. Liu, S. Schulte-Merker, J.P. Kanki, R. Plasterk, L.I. Zon, and A.T. Look, tp53 mutant zebrafish develop malignant peripheral nerve sheath tumors. Proc Natl Acad Sci U S A, 2005. 102(2): p. 407–12.PubMedCrossRefGoogle Scholar
  3. 3.
    Haramis, A.P., A. Hurlstone, Y. van der Velden, H. Begthel, M. van den Born, G.J. Offerhaus, and H.C. Clevers, Adenomatous polyposis coli-deficient zebrafish are susceptible to digestive tract neoplasia. EMBO Rep, 2006. 7(4): p. 444–9.PubMedGoogle Scholar
  4. 4.
    Langenau, D.M., D. Traver, A.A. Ferrando, J.L. Kutok, J.C. Aster, J.P. Kanki, S. Lin, E. Prochownik, N.S. Trede, L.I. Zon, and A.T. Look, Myc-induced T cell leukemia in transgenic zebrafish. Science, 2003. 299(5608): p. 887–90.PubMedCrossRefGoogle Scholar
  5. 5.
    Patton, E.E., H.R. Widlund, J.L. Kutok, K.R. Kopani, J.F. Amatruda, R.D. Murphey, S. Berghmans, E.A. Mayhall, D. Traver, C.D. Fletcher, J.C. Aster, S.R. Granter, A.T. Look, C. Lee, D.E. Fisher, and L.I. Zon, BRAF mutations are sufficient to promote nevi formation and cooperate with p53 in the genesis of melanoma. Curr Biol, 2005. 15(3): p. 249–54.PubMedCrossRefGoogle Scholar
  6. 6.
    Pardal, R., M.F. Clarke, and S.J. Morrison, Applying the principles of stem-cell biology to cancer. Nat Rev Cancer, 2003. 3(12): p. 895–902.PubMedCrossRefGoogle Scholar
  7. 7.
    Singh, S.K., I.D. Clarke, M. Terasaki, V.E. Bonn, C. Hawkins, J. Squire, and P.B. Dirks, Identification of a cancer stem cell in human brain tumors. Cancer Res, 2003. 63(18): p. 5821–8.Google Scholar
  8. 8.
    Al-Hajj, M., M.S. Wicha, A. Benito-Hernandez, S.J. Morrison, and M.F. Clarke, Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci U S A, 2003. 100(7): p. 3983–8.PubMedCrossRefGoogle Scholar
  9. 9.
    Bonnet, D. and J.E. Dick, Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med, 1997. 3(7): p. 730–7.PubMedCrossRefGoogle Scholar
  10. 10.
    Lapidot, T., C. Sirard, J. Vormoor, B. Murdoch, T. Hoang, J. Caceres-Cortes, M. Minden, B. Paterson, M.A. Caligiuri, and J.E. Dick, A cell initiating human acute myeloid leukaemia after transplantation into SCID mice. Nature, 1994. 367(6464): p. 645–8.PubMedCrossRefGoogle Scholar
  11. 11.
    Tan, B.T., C.Y. Park, L.E. Ailles, and I.L. Weissman, (2006) The cancer stem cell hypothesis: a work in progress. Lab Invest, 86(12): p. 1203–7.PubMedCrossRefGoogle Scholar
  12. 12.
    Traver, D., A. Winzeler, H.M. Stern, E.A. Mayhall, D.M. Langenau, J.L. Kutok, A.T. Look, and L.I. Zon, (2004) Effects of lethal irradiation in zebrafish and rescue by hemato-poietic cell transplantation. Blood, 104(5): p. 1298–305.PubMedCrossRefGoogle Scholar
  13. 13.
    Guryev, V., M.J. Koudijs, E. Berezikov, S.L. Johnson, R.H. Plasterk, F.J. van Eeden, and E. Cuppen,(2006) Genetic variation in the zebrafish. Genome Res, 16(4): p. 491–7.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Michael C. Dovey
    • 1
  • Leonard I. Zon
    • 2
  1. 1.Harvard Medical SchoolBostonUSA
  2. 2.Stem Cell Program and Division Hematology/Oncology Children’s Hospital and Dana Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Stem Cell InstituteHarvard Medical SchoolBostonUSA

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