Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Efficient tumour formation by single human melanoma cells

Nature volume 456, pages 593–598 (2008)Cite this article

Abstract

A fundamental question in cancer biology is whether cells with tumorigenic potential are common or rare within human cancers. Studies on diverse cancers, including melanoma, have indicated that only rare human cancer cells (0.1–0.0001%) form tumours when transplanted into non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice. However, the extent to which NOD/SCID mice underestimate the frequency of tumorigenic human cancer cells has been uncertain. Here we show that modified xenotransplantation assay conditions, including the use of more highly immunocompromised NOD/SCID interleukin-2 receptor gamma chain null (Il2rg-/-) mice, can increase the detection of tumorigenic melanoma cells by several orders of magnitude. In limiting dilution assays, approximately 25% of unselected melanoma cells from 12 different patients, including cells from primary and metastatic melanomas obtained directly from patients, formed tumours under these more permissive conditions. In single-cell transplants, an average of 27% of unselected melanoma cells from four different patients formed tumours. Modifications to xenotransplantation assays can therefore dramatically increase the detectable frequency of tumorigenic cells, demonstrating that they are common in some human cancers.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Get just this article for as long as you need it

$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Only rare human melanoma cells form tumours in NOD/SCID mice.
Figure 2: Modifications to the xenotransplantation assay reveal that many more human melanoma cells have tumorigenic potential than detected in NOD/SCID mice.
Figure 3: A high percentage of human melanoma cells are tumorigenic, but normal human cells are not.
Figure 4: Efficient tumour development from the xenotransplantation of single human melanoma cells.

References

  1. Schatton, T. et al. Identification of cells initiating human melanomas. Nature 451, 345–349 (2008)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  2. Li, C. et al. Identification of pancreatic cancer stem cells. Cancer Res. 67, 1030–1037 (2007)

    Article  CAS  PubMed  Google Scholar 

  3. Prince, M. E. et al. Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. Proc. Natl Acad. Sci. USA 104, 973–978 (2007)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  4. Wu, C. et al. Side population cells isolated from mesenchymal neoplasms have tumor initiating potential. Cancer Res. 67, 8216–8222 (2007)

    Article  CAS  PubMed  Google Scholar 

  5. Wang, J. C. et al. High level engraftment of NOD/SCID mice by primitive normal and leukemic hematopoietic cells from patients with chronic myeloid leukemia in chronic phase. Blood 91, 2406–2414 (1998)

    Article  CAS  PubMed  Google Scholar 

  6. Al-Hajj, M. et al. Prospective identification of tumorigenic breast cancer cells. Proc. Natl Acad. Sci. USA 100, 3983–3988 (2003)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  7. Cox, C. V. et al. Characterization of acute lymphoblastic leukemia progenitor cells. Blood 104, 2919–2925 (2004)

    Article  CAS  PubMed  Google Scholar 

  8. Hope, K. J., Jin, L. & Dick, J. E. Acute myeloid leukemia originates from a hierarchy of leukemic stem cell classes that differ in self-renewal capacity. Nature Immunol. 5, 738–743 (2004)

    Article  CAS  Google Scholar 

  9. Dalerba, P. et al. Phenotypic characterization of human colorectal cancer stem cells. Proc. Natl Acad. Sci. USA 104, 10158–10163 (2007)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  10. O’Brien, C. A. et al. A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 445, 106–110 (2007)

    Article  ADS  PubMed  CAS  Google Scholar 

  11. Ricci-Vitiani, L. et al. Identification and expansion of human colon-cancer-initiating cells. Nature 445, 111–115 (2007)

    Article  ADS  CAS  PubMed  Google Scholar 

  12. Williams, R. T., den Besten, W. & Sherr, C. J. Cytokine-dependent imatinib resistance in mouse BCR-ABL+, Arf-null lymphoblastic leukemia. Genes Dev. 21, 2283–2287 (2007)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Kelly, P. N. et al. Tumor growth need not be driven by rare cancer stem cells. Science 317, 337 (2007)

    Article  ADS  CAS  PubMed  Google Scholar 

  14. Somervaille, T. C. & Cleary, M. L. Identification and characterization of leukemia stem cells in murine MLL-AF9 acute myeloid leukemia. Cancer Cell 10, 257–268 (2006)

    Article  CAS  PubMed  Google Scholar 

  15. Agliano, A. et al. Human acute leukemia cells injected in NOD/LtSz-SCID/IL-γnull mice generate a faster and more efficient disease compared to other NOD/SCID-related strains. Int. J. Cancer 123, 2222–2227 (2008)

    Article  CAS  PubMed  Google Scholar 

  16. Feuring-Buske, M. et al. Improved engraftment of human acute myeloid leukemia progenitor cells in β-2-microglobulin-deficient NOD/SCID mice and in NOD/SCID mice transgenic for human growth factors. Leukemia 17, 760–763 (2003)

    Article  CAS  PubMed  Google Scholar 

  17. Kennedy, J. A. et al. Comment on ‘Tumor growth need not be driven by rare cancer stem cells’. Science 318, 1722; author reply 1722 (2007)

    Article  ADS  CAS  PubMed  Google Scholar 

  18. Pardal, R., Clarke, M. F. & Morrison, S. J. Applying the principles of stem-cell biology to cancer. Nature Rev. Cancer 3, 895–902 (2003)

    Article  CAS  Google Scholar 

  19. Wang, J. C. & Dick, J. E. Cancer stem cells: lessons from leukemia. Trends Cell Biol. 15, 494–501 (2005)

    Article  CAS  PubMed  Google Scholar 

  20. Shackleton, M. et al. Generation of a functional mammary gland from a single stem cell. Nature 439, 84–88 (2006)

    Article  ADS  CAS  PubMed  Google Scholar 

  21. Shultz, L. D. et al. Human lymphoid and myeloid cell development in NOD/LtSz-scid IL2Rγnull mice engrafted with mobilized human hemopoietic stem cells. J. Immunol. 174, 6477–6489 (2005)

    Article  CAS  PubMed  Google Scholar 

  22. Ito, M. et al. NOD/SCID/γc null mouse: an excellent recipient mouse model for engraftment of human cells. Blood 100, 3175–3182 (2002)

    Article  CAS  PubMed  Google Scholar 

  23. McKenzie, J. L. et al. Human short-term repopulating stem cells are efficiently detected following intrafemoral transplantation into NOD/SCID recipients depleted of CD122+ cells. Blood 106, 1259–1261 (2005)

    Article  CAS  PubMed  Google Scholar 

  24. Shultz, L. D. et al. Regulation of human short-term repopulating cell (STRC) engraftment in NOD/SCID mice by host CD122+ cells. Exp. Hematol. 31, 551–558 (2003)

    Article  PubMed  Google Scholar 

  25. Hamada, K. et al. Liver metastasis models of colon cancer for evaluation of drug efficacy using NOD/Shi-scid IL2Rγnull (NOG) mice. Int. J. Oncol. 32, 153–159 (2008)

    CAS  PubMed  Google Scholar 

  26. Suemizu, H. et al. Identification of a key molecular regulator of liver metastasis in human pancreatic carcinoma using a novel quantitative model of metastasis in NOD/SCID/γcnull (NOG) mice. Int. J. Oncol. 31, 741–751 (2007)

    CAS  PubMed  Google Scholar 

  27. Ishikawa, F. et al. Chemotherapy-resistant human AML stem cells home to and engraft within the bone-marrow endosteal region. Nature Biotechnol. 25, 1315–1321 (2007)

    Article  CAS  Google Scholar 

  28. Kleinman, H. K. & Martin, G. R. Matrigel: basement membrane matrix with biological activity. Semin. Cancer Biol. 15, 378–386 (2005)

    Article  CAS  PubMed  Google Scholar 

  29. Pretlow, T. G. et al. Transplantation of human prostatic carcinoma into nude mice in Matrigel. Cancer Res. 51, 3814–3817 (1991)

    CAS  PubMed  Google Scholar 

  30. Sweeney, T. M. et al. Basement membrane and the SIKVAV laminin-derived peptide promote tumor growth and metastases. Cancer Metastasis Rev. 10, 245–254 (1991)

    Article  CAS  PubMed  Google Scholar 

  31. Fridman, R. et al. Enhanced tumor growth of both primary and established human and murine tumor cells in athymic mice after coinjection with Matrigel. J. Natl Cancer Inst. 83, 769–774 (1991)

    Article  CAS  PubMed  Google Scholar 

  32. Singh, S. K. et al. Identification of human brain tumour initiating cells. Nature 432, 396–401 (2004)

    Article  ADS  CAS  PubMed  Google Scholar 

  33. Iwashita, T. et al. Hirschsprung disease is linked to defects in neural crest stem cell function. Science 301, 972–976 (2003)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  34. Bao, S. et al. Targeting cancer stem cells through L1CAM suppresses glioma growth. Cancer Res. 68, 6043–6048 (2008)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Frank, N. Y. et al. ABCB5-mediated doxorubicin transport and chemoresistance in human malignant melanoma. Cancer Res. 65, 4320–4333 (2005)

    Article  CAS  PubMed  Google Scholar 

  36. Yilmaz, O. H. et al. Pten dependence distinguishes haematopoietic stem cells from leukaemia-initiating cells. Nature 441, 475–482 (2006)

    Article  ADS  CAS  PubMed  Google Scholar 

  37. Horwich, A., Shipley, J. & Huddart, R. Testicular germ-cell cancer. Lancet 367, 754–765 (2006)

    Article  CAS  PubMed  Google Scholar 

  38. Kleinsmith, L. J. & Pierce, G. B. Multipotentiality of single embryonal carcinoma cells. Cancer Res. 24, 1544–1551 (1964)

    CAS  PubMed  Google Scholar 

  39. Bonnefoix, T. et al. Fitting limiting dilution experiments with generalized linear models results in a test of the single-hit Poisson assumption. J. Immunol. Methods 194, 113–119 (1996)

    Article  CAS  PubMed  Google Scholar 

  40. Nakajima, T. et al. Immunohistochemical demonstration of S100 protein in malignant melanoma and pigmented nevus, and its diagnostic application. Cancer 50, 912–918 (1982)

    Article  CAS  PubMed  Google Scholar 

  41. Fernandez, Y. et al. Differential regulation of noxa in normal melanocytes and melanoma cells by proteasome inhibition: therapeutic implications. Cancer Res. 65, 6294–6304 (2005)

    Article  CAS  PubMed  Google Scholar 

  42. Wang, Z. et al. Ablation of proliferating marrow with 5-fluorouracil allows partial purification of mesenchymal stem cells. Stem Cells 24, 1573–1582 (2006)

    Article  CAS  PubMed  Google Scholar 

  43. Bancroft, J. D. & Stevens, A. Theory and Practice of Histological Techniques (Churchill Livingstone, 1990)

    Google Scholar 

Download references

Acknowledgements

This work was supported by the Howard Hughes Medical Institute and by the Allen H. Blondy Research Fellowship. The University of Michigan Melanoma Bank was supported by a gift from Lewis and Lillian Becker. Flow cytometry was partly supported by the University of Michigan Comprehensive Cancer Center grant from the National Institutes of Health CA46592. We thank: D. Adams, M. White and the University of Michigan Flow Cytometry Core Facility for support; N. McAnsh and the University of Michigan Cancer Centre Histology Core for histological studies; G. K. Smyth for assistance with statistics; and Z. Azizan for support with tissue collection. Antibody production was supported in part by the National Institute of Diabetes, Digestive, and Kidney Diseases, grant NIH5P60-DK20572 to the Michigan Diabetes Research and Training Center. Some antibodies were provided by Caltag or by eBioscience to screen for cancer stem-cell markers. Human primary melanocyte cultures were provided by M. Soengas. Human mesenchymal stem cells were provided by Z. Wang and P. Krebsbach. E.Q. was supported by the Spanish Ministry of Education and the Marie Curie Outgoing International Fellowship from the European Commission. M.S. was supported by the Australian National Health and Medical Research Council, the Human Frontiers Science Program and Australia Post.

Author Contributions E.Q., M.S. and S.J.M. planned the project. E.Q. and M.S. performed the experiments, and analysed data with S.J.M. M.S.S. and T.M.J. obtained consent from the patients and surgically obtained many of the melanoma specimens. D.R.F. performed all pathology and diagnosed the tumours with T.M.J. T.M.J. banked the melanomas, and provided clinical information. E.Q., M.S. and S.J.M. wrote the paper.

Author information

Author notes

  1. Elsa Quintana and Mark Shackleton: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Internal Medicine, Howard Hughes Medical Institute, Life Sciences Institute, and Center for Stem Cell Biology, University of Michigan, Ann Arbor, Michigan 48109-2216, USA,

    Elsa Quintana, Mark Shackleton & Sean J. Morrison

  2. Department of Dermatology, University of Michigan, Ann Arbor, Michigan 48109, USA,

    Timothy M. Johnson

Authors
  1. Elsa Quintana
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mark Shackleton
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michael S. Sabel
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Douglas R. Fullen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Timothy M. Johnson
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sean J. Morrison
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sean J. Morrison.

Supplementary information

Supplementary Information

This file contains Supplementary Tables 1, 2 and Supplementary Figures 1-6 with legends. (PDF 3549 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Quintana, E., Shackleton, M., Sabel, M. et al. Efficient tumour formation by single human melanoma cells. Nature 456, 593–598 (2008). https://doi.org/10.1038/nature07567

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature07567

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing