Summary
The study of key mechanisms and molecules involved in the regulation of hematopoiesis in mouse models has been greatly facilitated by multi-parameter flow cytometry. Subpopulations of hematopoietic stem and progenitor cells can be identified and characterized using this technique. Furthermore, fluorescence-activated cell sorting (FACS) can prospectively isolate functionally-defined subpopulations of hematopoietic cells for use in further in vitro or in vivo analysis. The chapter describes methodology for preparing samples from mouse bone marrow, staining cells with fluorochrome conjugated antibodies, enrichment of HSC and progenitors, and finally data analysis.
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References
Kondo M, Weissman IL, Akashi K. (1997). Identifi-cation of clonogenic common lymphoid progenitors in mouse bone marrow. Cell; 91(5): 661–72.
Akashi K, Traver D, Miyamoto T, Weissman IL. (2000). A clonogenic common myeloid progenitor that gives rise to all myeloid lineages. Nature; 404(6774): 193–7.
So CW, Karsunky H, Wong P, Weissman IL, Cleary ML. (2004). Leukemic transformation of hematopoietic progenitors by MLL-GAS7 in the absence of Hoxa7 or Hoxa9. Blood; 103(8): 3192–9.
So CW, Karsunky H, Passegue E, Cozzio A, Weissman IL, Cleary ML. (2003). MLL-GAS7 transforms multipotent hematopoietic progenitors and induces mixed lineage leukemias in mice. Cancer Cell; 3(2): 161–71.
Cozzio A, Passegue E, Ayton PM, Karsunky H, Cleary ML, Weissman IL. (2003). Similar MLL-associated leukemias arising from self-renewing stem cells and short-lived myeloid progenitors. Genes Dev; 17(24): 3029–35.
Christensen JL, Weissman IL. (2001). Flk-2 is a marker in hematopoietic stem cell differentiation: a simple method to isolate long-term stem cells. Proc Natl Acad Sci U S A; 98(25): 14541–6.
Adolfsson J, Borge OJ, Bryder D, et al. (2001). Upregulation of Flt3 expression within the bone marrow Lin(−)Sca1(+)c-kit(+) stem cell compartment is accompanied by loss of self-renewal capacity. Immunity; 15(4): 659–69.
Yang L, Bryder D, Adolfsson J, et al. (2005). Identification of Lin-Sca1+kit+CD34+Flt3- short-term hematopoietic stem cells capable of rapidly reconstituting and rescuing myeloablated transplant recipients. Blood; 105(7): 2717–23.
Morrison SJ, Weissman IL. (1994). The long-term repopulating subset of hematopoietic stem cells is deterministic and isolatable by phenotype. Immunity; 1(8): 661–73.
Morrison SJ, Wandycz AM, Hemmati HD, Wright DE, Weissman IL. (1997). Identification of a lineage of multipotent hematopoietic progenitors. Development; 124(10): 1929–39.
Kiel MJ, Yilmaz OH, Iwashita T, Yilmaz OH, Terhorst C, Morrison SJ. (2005). SLAM family receptors distinguish hematopoietic stem and progenitor cells and reveal endothelial niches for stem cells. Cell; 121(7): 1109.
Ikuta K, Weissman IL. (1992). Evidence that hematopoietic stem cells express mouse c-kit but do not depend on steel factor for their generation. Proc Natl Acad Sci; 89(4): 1502–6.
Spangrude GJ, Heimfeld S, Weissman IL. (1988). Purification and characterization of mouse hematopoietic stem cells. Science; 241(4861): 58–62.
Li CL, Johnson GR. (1995). Murine hematopoietic stem and progenitor cells: I. Enrichment and biologic characterization. Blood; 85(6): 1472–9.
Acknowledgments
The authors thank Ian Titley and members of the So’s lab for constructive advice. The Association for International Cancer Research (AICR), Cancer Research UK, the Kay Kendall Leukaemia Fund, Medical Research Council, Wellcome Trust, and Leukaemia Research Fund supported this work. Eric So is an AICR fellow and an EMBO young investigator.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Yeung, J., Wai Eric So, a. (2009). Identification and Characterization of Hematopoietic Stem and Progenitor Cell Populations in Mouse Bone Marrow by Flow Cytometry. In: Eric So, C.W. (eds) Leukemia. Methods in Molecular Biology™, vol 538. Humana Press. https://doi.org/10.1007/978-1-59745-418-6_15
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DOI: https://doi.org/10.1007/978-1-59745-418-6_15
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