Analysis of Bcr-Abl Function Using an In Vitro Embryonic Stem Cell Differentiation System

  • Takumi Era
  • Stephane Wong
  • Owen N. Witte
Part of the Methods in Molecular Biology™ book series (MIMB, volume 185)


Numerous causative genes involved in human cancers, such as leukemias and lymphomas, have been identified (1). Considerable evidence has accumulated showing that disruptions of these genes can affect hematopoietic cell differentiation and growth. However, it remains largely unknown how these molecules function directly in hematopoietic stem cells.


Differentiation Culture Medium Leukemia Inhibitory Factor Resuspend Cell Hematopoietic Development Liquid Nitrogen Tank 
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  1. 1.
    Lowenberg, B., Downing, J. R., and Burnett, A. (1999) Acute myeloid leukemia. N. Engl. J. Med. 341, 1051–1062.PubMedCrossRefGoogle Scholar
  2. 2.
    Sawyers, C. L. (1999) Chronic myeloid leukemia. N. Engl. J. Med. 340, 1330–1340.PubMedCrossRefGoogle Scholar
  3. 3.
    Groffen, J., Stephenson, J. R., Heisterkamp, N., de Klein, A., Bartram, C. R., and Grosveld, G. (1984) Philadelphia chromosomal breakpoints are clustered within a limited region, bcr, on chromosome 22. Cell 36, 93–99.PubMedCrossRefGoogle Scholar
  4. 4.
    Konopka, J. B., Watanabe, S. M., and Witte, O. N. (1984) An alteration of the human c-abl protein in K562 leukemia cells unmasks associated tyrosine kinase activity. Cell 37, 1035–1042.PubMedCrossRefGoogle Scholar
  5. 5.
    Shtivelman, E., Lifshitz, B., Gale, R. P., Roe, B. A., and Canaani, E. (1985) Fused transcript of abl and bcr genes in chronic myelogenous leukaemia. Nature 315, 550–554.PubMedCrossRefGoogle Scholar
  6. 6.
    McLaughlin, J., Chianese, E., and Witte, O. N. (1987) In vitro transformation of immature hematopoietic cells by the P210 BCR/Abl oncogene product of the Philadelphia chromosome. Proc. Natl. Acad. Sci. USA 84, 6558–6562.PubMedCrossRefGoogle Scholar
  7. 7.
    Lugo, T. G., Pendergast, A., Muller, A. J., and Witte, O. N. (1990) Tyrosine kinase activity and transformation potency of bcr-abl oncogene products. Science 247, 1079–1082.PubMedCrossRefGoogle Scholar
  8. 8.
    Daley, G. Q. and Baltimore, D. (1988) Transformation of an interleukin 3-dependent hematopoietic cell line by the chronic myelogenous leukemia-specific P210+bcr/abl~ protein. Proc. Natl. Acad. Sci. USA 85, 9312–93PubMedCrossRefGoogle Scholar
  9. 9.
    Gishizky, M. L. and Witte, O. N. (1992) Initiation of deregulated growth of multipotent progenitor cells by bcr-abl in vitro. Science 256, 836–839.PubMedCrossRefGoogle Scholar
  10. 10.
    Daley, G. Q., Van Etten, R. A., and Baltimore, D. (1990) Induction of chronic myelogenous leukemia in mice by the P210bcr/abl gene of the Philadelphia chromosome. Science 247, 824–830.PubMedCrossRefGoogle Scholar
  11. 11.
    Kelliher, M. A., McLaughlin, J., Witte, O. N., and Rosenberg, N. (1990) Induction of a chronic myelogenous leukemia-like syndrome in mice with v-abl and BCR/ABL. Proc. Natl. Acad. Sci. USA 87, 6649–6653.PubMedCrossRefGoogle Scholar
  12. 12.
    Honda, H., Oda, H., Suzuki, T., Takahashi, T., Witte, O. N., Ozawa, K., et al. (1998) Development of acute lymphoblastic leukemia and myeloproliferative disorder in transgenic mice expressing p210bcr/abl: a novel transgenic model for human Ph1-positive leukemias. Blood 91, 2067–2075.PubMedGoogle Scholar
  13. 13.
    Pear, W. S., Miller, J. P., Xu, L., Pui, J. C., Soffer, B., Quackenbush, R. C., et al. (1998) Efficient and rapid induction of a chronic myelogenous leukemia-like myeloproliferative disease in mice receiving P210 bcr/abl-transduced bone marrow. Blood 92, 3780–3792.PubMedGoogle Scholar
  14. 14.
    Nakano, T., Kodama, H., and Honjo, T. (1994) Generation of lymphohematopoietic cells from embryonic stem cells in culture. Science 265, 1098–1101.PubMedCrossRefGoogle Scholar
  15. 15.
    Nakano, T. and Kodama, H. H. (1996) In vitro development of primitive and definitive erythrocytes from different precursors. Science 272, 722–724.PubMedCrossRefGoogle Scholar
  16. 16.
    Era, T., Takahashi, T., Sakai, K., Kawamura, K., and Nakano, T. (1997) Thrombopoietin enhances proliferation and differentiation of murine yolk sac erythroid progenitors. Blood 89, 1207–1213.PubMedGoogle Scholar
  17. 17.
    Era, T., Takagi, T., Takahashi, T., Bories, J. C., and Nakano, T. (2000) Characterization of hematopoietic lineage-specific gene expression by ES cell in vitro differentiation induction system. Blood 95, 870–878.PubMedGoogle Scholar
  18. 18.
    Gossen, M. and Bujard, H. (1992) Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. USA 89, 5547–5551.PubMedCrossRefGoogle Scholar
  19. 19.
    Era, T. and Witte, O. N. (2000) Regulated expression of P210 Bcr-Abl during embryonic stem cell differentiation stimulates multipotential progenitor expansion and myeloid cell fate. Proc. Natl. Acad. Sci. USA 97, 1737–1742.PubMedCrossRefGoogle Scholar
  20. 20.
    Baron, U., Gossen, M., and Bujard, H. (1997) Tetracycline-controlled transcription in eukaryotes: novel transactivators with graded transactivation potential. Nucleic Acids Res. 25, 2723–2729.PubMedCrossRefGoogle Scholar
  21. 21.
    Niwa, H., Burdon, T., Chambers, I., and Smith, A. (1998) Self-renewal of pluripotent embryonic stem cells is mediated via activation of STAT3. Genes Dev. 12, 2048–2060.PubMedCrossRefGoogle Scholar
  22. 22.
    Niwa, H., Yamamura, K., and Miyazaki, J. (1991) Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene 108, 193–199.PubMedCrossRefGoogle Scholar
  23. 23.
    Robertson, E. J. (1987) Embryo-derived stem cells, in Teratocacinomas and Embryonic Stem Cell: A Practical Approach (Robertson, E. J., ed.), IRL Press, New York, pp. 71–112.Google Scholar
  24. 24.
    Wurst, W. and Joyner, A. L. (1993) Gene Targeting: A Practical Approach, vol. 126. IRL Press, New York.Google Scholar
  25. 25.
    Chambard, J. C. and Pognonec, P. (1998) A reliable way of obtaining stable inducible clones. Nucleic Acids Res. 26, 3443–3444.PubMedCrossRefGoogle Scholar
  26. 26.
    Ramirez-Solis, R., Davis, A. C., and Bradley, A. (1993) Gene targeting in embryonic stem cells. Methods Enzymol. 225, 855–878.PubMedCrossRefGoogle Scholar
  27. 27.
    Cho, S. K., Webber, T. D., Carlyle, J. R., Nakano, T., Lewis, S. M., and Zuniga-Pflucker, J. C. (1999) Functional characterization of B lymphocytes generated in vitro from embryonic stem cells. Proc. Natl. Acad. Sci. USA 96, 9797–9802.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Takumi Era
    • 1
  • Stephane Wong
    • 1
  • Owen N. Witte
    • 1
  1. 1.Howard Hughes Medical InstituteUniversity of CaliforniaLos Angeles

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