Letter | Published:

Identification of human brain tumour initiating cells

Naturevolume 432pages396401 (2004) | Download Citation



The cancer stem cell (CSC) hypothesis suggests that neoplastic clones are maintained exclusively by a rare fraction of cells with stem cell properties1,2. Although the existence of CSCs in human leukaemia is established3,4, little evidence exists for CSCs in solid tumours, except for breast cancer5. Recently, we prospectively isolated a CD133+ cell subpopulation from human brain tumours that exhibited stem cell properties in vitro6. However, the true measures of CSCs are their capacity for self renewal and exact recapitulation of the original tumour1,2,7. Here we report the development of a xenograft assay that identified human brain tumour initiating cells that initiate tumours in vivo. Only the CD133+ brain tumour fraction contains cells that are capable of tumour initiation in NOD-SCID (non-obese diabetic, severe combined immunodeficient) mouse brains. Injection of as few as 100 CD133+ cells produced a tumour that could be serially transplanted and was a phenocopy of the patient's original tumour, whereas injection of 105 CD133- cells engrafted but did not cause a tumour. Thus, the identification of brain tumour initiating cells provides insights into human brain tumour pathogenesis, giving strong support for the CSC hypothesis as the basis for many solid tumours5, and establishes a previously unidentified cellular target for more effective cancer therapies.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1

    Reya, T., Morrison, S. J., Clarke, M. F. & Weissman, I. L. Stem cells, cancer, and cancer stem cells. Nature 414, 105–111 (2001)

  2. 2

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

  3. 3

    Lapidot, T. et al. A cell initiating human acute myeloid leukaemia after transplantation into SCID mice. Nature 367, 645–648 (1994)

  4. 4

    Bonnet, D. & Dick, J. E. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nature Med. 3, 730–737 (1997)

  5. 5

    Al-Hajj, M., Wicha, M. S., Benito-Hernandez, A., Morrison, S. J. & Clarke, M. F. Prospective identification of tumorigenic breast cancer cells. Proc. Natl Acad. Sci. USA 100, 3983–3988 (2003)

  6. 6

    Singh, S. K. et al. Identification of a cancer stem cell in human brain tumors. Cancer Res. 63, 5821–5828 (2003)

  7. 7

    Dick, J. E. Breast cancer stem cells revealed. Proc. Natl Acad. Sci. USA 100, 3547–3549 (2003)

  8. 8

    Ignatova, T. N. et al. Human cortical glial tumors contain neural stem-like cells expressing astroglial and neuronal markers in vitro. Glia 39, 193–206 (2002)

  9. 9

    Hemmati, H. D. et al. Cancerous stem cells can arise from pediatric brain tumors. Proc. Natl Acad. Sci. USA 100, 15178–15183 (2003)

  10. 10

    Kondo, T., Setoguchi, T. & Taga, T. Persistence of a small subpopulation of cancer stem-like cells in the C6 glioma cell line. Proc. Natl Acad. Sci. USA 101, 781–786 (2004)

  11. 11

    Miraglia, S. et al. A novel five-transmembrane hematopoietic stem cell antigen: isolation, characterization, and molecular cloning. Blood 90, 5013–5021 (1997)

  12. 12

    Corbeil, D., Roper, K., Weigmann, A. & Huttner, W. B. AC133 hematopoietic stem cell antigen: human homologue of mouse kidney prominin or distinct member of a novel protein family? Blood 91, 2625–2626 (1998)

  13. 13

    Uchida, N. et al. Direct isolation of human central nervous system stem cells. Proc. Natl Acad. Sci. USA 97, 14720–14725 (2000)

  14. 14

    Houchens, D. P., Ovejera, A. A., Riblet, S. M. & Slagel, D. E. Human brain tumor xenografts in nude mice as a chemotherapy model. Eur. J. Cancer Clin. Oncol. 19, 799–805 (1983)

  15. 15

    Hu, B. et al. Angiopoietin-2 induces human glioma invasion through the activation of matrix metalloprotease-2. Proc. Natl Acad. Sci. USA 100, 8904–8909 (2003)

  16. 16

    Russell, D. & Rubenstein, L. Pathology of Tumors of the Central Nervous System (Williams and Wilkins, Baltimore, 1989)

  17. 17

    Lendahl, U., Zimmerman, L. B. & McKay, R. D. CNS stem cells express a new class of intermediate filament protein. Cell 60, 585–595 (1990)

  18. 18

    Kleihues, P. et al. The WHO classification of tumors of the nervous system. J. Neuropathol. Exp. Neurol. 61, 215–225; discussion 226–229 (2002)

  19. 19

    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)

  20. 20

    Cozzio, A. et al. Similar MLL-associated leukemias arising from self-renewing stem cells and short-lived myeloid progenitors. Genes Dev. 17, 3029–3035 (2003)

  21. 21

    Passegue, E., Jamieson, C. H., Ailles, L. E. & Weissman, I. L. Normal and leukemic hematopoiesis: are leukemias a stem cell disorder or a reacquisition of stem cell characteristics? Proc. Natl Acad. Sci. USA 100(suppl. 1), 11842–11849 (2003)

  22. 22

    Holland, E. C., Hively, W. P., DePinho, R. A. & Varmus, H. E. A constitutively active epidermal growth factor receptor cooperates with disruption of G1 cell-cycle arrest pathways to induce glioma-like lesions in mice. Genes Dev. 12, 3675–3685 (1998)

  23. 23

    Holland, E. C. Gliomagenesis: genetic alterations and mouse models. Nature Rev. Genet. 2, 120–129 (2001)

  24. 24

    Bachoo, R. M. et al. Epidermal growth factor receptor and Ink4a/Arf: convergent mechanisms governing terminal differentiation and transformation along the neural stem cell to astrocyte axis. Cancer Cell 1, 269–277 (2002)

  25. 25

    Oliver, T. G. & Wechsler-Reya, R. J. Getting at the root and stem of brain tumors. Neuron 42, 885–888 (2004)

  26. 26

    Potgens, A. J., Bolte, M., Huppertz, B., Kaufmann, P. & Frank, H. G. Human trophoblast contains an intracellular protein reactive with an antibody against CD133–a novel marker for trophoblast. Placenta 22, 639–645 (2001)

  27. 27

    Bayani, J. et al. Molecular cytogenetic analysis of medulloblastomas and supratentorial primitive neuroectodermal tumors by using conventional banding, comparative genomic hybridization, and spectral karyotyping. J. Neurosurg. 93, 437–448 (2000)

  28. 28

    Galli, R. et al. Isolation and characterization of tumorigenic, stem-like neural precursors from human glioblastoma. Cancer Res. 64, 7011–7021 (2004)

Download references


We thank M. Borden, J. Ma, I. Diplock, M. Ho and C. Gibson for technical assistance, and we are grateful to V. Bonn, L. Davidson, N. Lifshitz and J. Chen of the Mouse Imaging Centre for help with neuroimaging. We thank J. Dick for discussions. S. Singh was supported by a a Terry Fox Foundation fellowship from the Canadian Cancer Society, the Neurosurgical Research and Education Foundation and the American Brain Tumor Association. This work was supported by the Canadian Cancer Society, the Canadian Institutes of Health Research, the Foundation of The Hospital for Sick Children, BrainChild and the Jack Baker and Jessica Durigon family funds.

Author information


  1. The Arthur and Sonia Labatt Brain Tumor Research Centre, University of Toronto, 555 University Avenue, Toronto, M5G 1X8, Canada

    • Sheila K. Singh
    • , Cynthia Hawkins
    • , Ian D. Clarke
    • , Takuichiro Hide
    •  & Peter B. Dirks
  2. Program in Developmental Biology, University of Toronto, 555 University Avenue, Toronto, M5G 1X8, Canada

    • Sheila K. Singh
    • , Ian D. Clarke
    • , Takuichiro Hide
    •  & Peter B. Dirks
  3. Division of Neurosurgery, University of Toronto, 555 University Avenue, M5G 1X8, Toronto, Canada

    • Sheila K. Singh
    • , Michael D. Cusimano
    •  & Peter B. Dirks
  4. Department of Pediatric Laboratory Medicine, University of Toronto, 555 University Avenue, M5G 1X8, Toronto, Canada

    • Cynthia Hawkins
  5. Integrative Biology Program, The Hospital for Sick Children and University of Toronto, 555 University Avenue, M5G 1X8, Toronto, Canada

    • R. Mark Henkelman
  6. Ontario Cancer Institute and University of Toronto, 610 University Avenue, M5G 2M9, Toronto, Canada

    • Jeremy A. Squire
    •  & Jane Bayani
  7. Division of Neurosurgery, St Michael's Hospital and University of Toronto, 30 Bond Street, M5B 1W8, Toronto, Canada

    • Michael D. Cusimano


  1. Search for Sheila K. Singh in:

  2. Search for Cynthia Hawkins in:

  3. Search for Ian D. Clarke in:

  4. Search for Jeremy A. Squire in:

  5. Search for Jane Bayani in:

  6. Search for Takuichiro Hide in:

  7. Search for R. Mark Henkelman in:

  8. Search for Michael D. Cusimano in:

  9. Search for Peter B. Dirks in:

Competing interests

The authors declare that they have no competing financial interests.

Corresponding author

Correspondence to Peter B. Dirks.

Supplementary information

  1. Supplementary Table 1

    Patient information. (DOC 23 kb)

  2. Supplementary Table 2

    CD133 frequency and primary sphere formation rate of sorted tumours. (DOC 24 kb)

  3. Supplementary Table 3

    Summary of tumour cell injections into NOD–SCID mice. (DOC 28 kb)

  4. Supplementary Table 4

    Comparison of histology and immunohistochemistry of CD133+ xenotransplanted tumours and original patient tumours. (DOC 39 kb)

  5. Supplementary Figure 1

    Phenotypic characteristics of glioblastoma and medulloblastoma xenografts. (DOC 273 kb)

  6. Supplementary Figure 2

    CD133+ and CD133- cells from an original patient medulloblastoma show clonal karyotype abnormalities. (DOC 248 kb)

  7. Supplementary Figure 3

    CD133+ and CD133- cells from patient GBMs show clonal karyotype abnormalities. (DOC 730 kb)

  8. Supplementary Figure 4

    EGFR Amplification in GBM and resulting xenografts. (DOC 758 kb)

About this article

Publication history



Issue Date



Further reading


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.