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.
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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.
The authors declare that they have no competing financial interests.
Patient information. (DOC 23 kb)
CD133 frequency and primary sphere formation rate of sorted tumours. (DOC 24 kb)
Summary of tumour cell injections into NOD–SCID mice. (DOC 28 kb)
Comparison of histology and immunohistochemistry of CD133+ xenotransplanted tumours and original patient tumours. (DOC 39 kb)
Phenotypic characteristics of glioblastoma and medulloblastoma xenografts. (DOC 273 kb)
CD133+ and CD133- cells from an original patient medulloblastoma show clonal karyotype abnormalities. (DOC 248 kb)
CD133+ and CD133- cells from patient GBMs show clonal karyotype abnormalities. (DOC 730 kb)
EGFR Amplification in GBM and resulting xenografts. (DOC 758 kb)
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Singh, S., Hawkins, C., Clarke, I. et al. Identification of human brain tumour initiating cells. Nature 432, 396–401 (2004). https://doi.org/10.1038/nature03128
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