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Identification of Murine and Human Acute Myeloid Leukemia Stem Cells

  • Aniruddha J. Deshpande
  • Farid Ahmed
  • Christian Buske
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 568)

Summary

There is now compelling evidence to show that tumors, once believed to be a homogeneous mass of abnormally proliferative cells, comprise a heterogeneous population of transformed cells resembling the hierarchically organized populations in the corresponding tissue. At the top of this tumor hierarchy are the cancer stem cells (CSCs) which are critical for tumor growth and maintenance as they constantly replenish the tumor bulk and are believed to be resistant to most conventional chemotherapies. The first evidence for both malignant hierarchy and CSCs came from studies on acute myeloid leukemia (AML) (1, 2) followed by mounting evidence in other types of cancer ( 3, 4, 5 ) . Murine models of leukemia have been the proving ground for the elucidation of key novel concepts in this nascent field of CSC research. A spate of recent studies highlighting the importance of CSCs in cancer has accentuated the need for identifying and characterizing these cells. The frequency of CSCs in the leukemic bulk (LSCs) is usually estimated by in vivo limiting-dilution transplantation assays of leukemic cells into recipient animal hosts in which identical leukemias can be regenerated. Each cell that is capable of propagating the leukemia in secondary recipients is termed a leukemia-propagating cell (LPC).

LSC candidates have been typically identified by the systematic dissection of compartments within a heterogeneous tumor to determine the subpopulation which shows maximal enrichment for LPCs. This is performed by determining the frequency of LPCs in each of those purified subpopulations, as has been described in both human and murine models of leukemia ( 1, 2, 6 ). It is important to mention that even though leukemia repopulation is often used as a surrogate for LSC estimation, the term LPCs can be equated with LSCs only if the phenotypic heterogeneity of the original tumor mass is regenerated upon tumor propagation into secondary recipients.

The first part of the chapter deals with the estimation of LPCs in syngenic or congenic murine–murine models. In the second part, murine xenograft models for the estimation of LSCs in human leukemias are described.

Key words

AML Cancer stem cells Leukemia Limiting dilution assays Leukemic stem cells Mouse bone marrow transplantation 

References

  1. 1.
    Bonnet, D. and J.E. Dick (1997), Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med. 3(7): p. 730–7.PubMedCrossRefGoogle Scholar
  2. 2.
    Lapidot, T., et al. (1994), A cell initiating human acute myeloid leukaemia after transplantation into SCID mice. Nature. 367(6464): p. 645–8.PubMedCrossRefGoogle Scholar
  3. 3.
    Al-Hajj, M., et al. (2003), Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci U S A. 100(7): p. 3983–8.PubMedCrossRefGoogle Scholar
  4. 4.
    Singh, S.K., et al. (2004), Identification of human brain tumour initiating cells. Nature. 432(7015): 396–401.PubMedCrossRefGoogle Scholar
  5. 5.
    O’Brien, C.A., et al. (2007), A human colon cancer cell capable of initiating tumour growth in immuno-deficient mice. Nature. 445(7123): p. 106–10.PubMedCrossRefGoogle Scholar
  6. 6.
    Deshpande, A.J., et al. (2006), Acute myeloid leukemia is propagated by a leukemic stem cell with lymphoid characteristics in a mouse model of CALM/AF10-positive leukemia. Cancer Cell. 10(5): p. 363–74.PubMedCrossRefGoogle Scholar
  7. 7.
    Szilvassy, S.J., et al. (1990), Quantitative assay for totipotent reconstituting hematopoietic stem cells by a competitive repopulation strategy. Proc Natl Acad Sci U S A. 87(22): p. 8736–40.PubMedCrossRefGoogle Scholar
  8. 8.
    Cozzio, A., et al. (2003), Similar MLL-associated leukemias arising from self-renewing stem cells and short-lived myeloid progenitors. Genes Dev. 17(24): p. 3029–35.PubMedCrossRefGoogle Scholar
  9. 9.
    So, C.W., et al. (2003), MLL-GAS7 transforms multipotent hematopoietic progenitors and induces mixed lineage leukemias in mice. Cancer Cell. 3(2): p. 161–71.PubMedCrossRefGoogle Scholar
  10. 10.
    Huntly, B.J., et al. (2004), MOZ-TIF2, but not BCR-ABL, confers properties of leukemic stem cells to committed murine hematopoietic progenitors. Cancer Cell. 6(6): p. 587–96.PubMedCrossRefGoogle Scholar
  11. 11.
    Ailles, L.E., B. Gerhard, and D.E. Hogge (1997), Detection and characterization of primitive malignant and normal progenitors in patients with acute myelogenous leukemia using long-term coculture with supportive feeder layers and cytokines. Blood. 90(7): p. 2555–64.PubMedGoogle Scholar
  12. 12.
    Buick, R.N., J.E. Till, and E.A. McCulloch (1977), Colony assay for proliferative blast cells circulating in myeloblastic leukaemia. Lancet. 1(8016): p. 862–3.PubMedCrossRefGoogle Scholar
  13. 13.
    Sutherland, H.J., et al. (1991), Differential regulation of primitive human hematopoietic cells in long-term cultures maintained on genetically engineered murine stromal cells. Blood. 78(3): p. 666–72.Google Scholar
  14. 14.
    Ailles, L.E., et al. (1999), Growth characteristics of acute myelogenous leukemia progenitors that initiate malignant hematopoiesis in nonobese diabetic/severe combined immunodeficient mice. Blood. 94(5): p. 1761–72.PubMedGoogle Scholar
  15. 15.
    Pearce, D.J., et al. (2006), AML engraftment in the NOD/SCID assay reflects the outcome of AML: implications for our understanding of the heterogeneity of AML. Blood. 107(3): p. 1166–73.PubMedCrossRefGoogle Scholar
  16. 16.
    Feuring-Buske, M., et al. (2002), A diphtheria toxin-interleukin 3 fusion protein is cytotoxic to primitive acute myeloid leukemia progenitors but spares normal progenitors. Cancer Res. 62(6): p. 1730–6.PubMedGoogle Scholar
  17. 17.
    Terpstra, W., et al. (1997), Diphtheria toxin fused to granulocyte-macrophage colony-stimulating factor eliminates acute myeloid leukemia cells with the potential to initiate leukemia in immunodeficient mice, but spares normal hemopoietic stem cells. Blood. 90(9): p. 3735–42.PubMedGoogle Scholar
  18. 18.
    Hosen, N., et al. (2007), CD96 is a leukemic stem cell-specific marker in human acute myeloid leukemia. Proc Natl Acad Sci U S A. 104(26): p. 11008–13.PubMedCrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Aniruddha J. Deshpande
    • 1
  • Farid Ahmed
    • 1
  • Christian Buske
    • 1
  1. 1.GSF National Research Center for Environment and Health and the Clinical Cooperative Group – Leukemia, Department of Medicine IIIKlinikum GrosshadernMunichGermany

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