Application of polychromatic flow cytometry to identify novel subsets of circulating cells with angiogenic potential

Authors

  • Myka L. Estes,

    1. Department of Pediatrics, Indiana University, Indianapolis, Indiana
    2. Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University, Indianapolis, Indiana
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    • The first two authors contributed equally to this work.

  • Julie A. Mund,

    1. Department of Pediatrics, Indiana University, Indianapolis, Indiana
    2. Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University, Indianapolis, Indiana
    3. Indiana University Melvin and Bren Simon Cancer Center, Department of Hematology/Oncology, Indianapolis, Indiana
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    • The first two authors contributed equally to this work.

  • Laura E. Mead,

    1. Department of Pediatrics, Indiana University, Indianapolis, Indiana
    2. Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University, Indianapolis, Indiana
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  • Daniel N. Prater,

    1. Department of Pediatrics, Indiana University, Indianapolis, Indiana
    2. Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University, Indianapolis, Indiana
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  • Shanbao Cai,

    1. Department of Pediatrics, Indiana University, Indianapolis, Indiana
    2. Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University, Indianapolis, Indiana
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  • Haiyan Wang,

    1. Department of Pediatrics, Indiana University, Indianapolis, Indiana
    2. Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University, Indianapolis, Indiana
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  • Karen E. Pollok,

    1. Department of Pediatrics, Indiana University, Indianapolis, Indiana
    2. Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University, Indianapolis, Indiana
    3. Indiana University Melvin and Bren Simon Cancer Center, Department of Hematology/Oncology, Indianapolis, Indiana
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  • Michael P. Murphy,

    1. Department of Surgery, Clarian Health Partners, IU/Methodist/Riley, Indianapolis, Indiana
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  • Caroline S. T. An,

    1. Department of Pathology, Clarian Health Partners, IU/Methodist/Riley, Indianapolis, Indiana
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  • Edward F. Srour,

    1. Department of Pediatrics, Indiana University, Indianapolis, Indiana
    2. Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University, Indianapolis, Indiana
    3. Indiana University Melvin and Bren Simon Cancer Center, Department of Hematology/Oncology, Indianapolis, Indiana
    4. Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana
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  • David A. Ingram Jr.,

    Corresponding author
    1. Department of Pediatrics, Indiana University, Indianapolis, Indiana
    2. Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University, Indianapolis, Indiana
    3. Indiana University Melvin and Bren Simon Cancer Center, Department of Hematology/Oncology, Indianapolis, Indiana
    4. Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana
    • Department of Pediatrics, Indiana University School of Medicine, 1044 West Walnut St, R4-470, Indianapolis, IN, 46202, USA
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  • Jamie Case

    Corresponding author
    1. Department of Pediatrics, Indiana University, Indianapolis, Indiana
    2. Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University, Indianapolis, Indiana
    3. Indiana University Melvin and Bren Simon Cancer Center, Department of Hematology/Oncology, Indianapolis, Indiana
    • Department of Pediatrics, Indiana University School of Medicine, 1044 West Walnut St, R4-470, Indianapolis, IN, 46202, USA
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Abstract

Defining whether human circulating proangiogenic cells represent a subset of the hematopoietic system and express CD45 or are hematopoietic derivatives that do not express CD45 (and are called endothelial progenitor cells) remains controversial. We have previously developed a polychromatic flow cytometry (PFC) protocol to isolate subsets of hematopoietic cells and we now identify the circulating pool of CD34+CD45dim cells representing functional circulating hematopoietic stem and progenitor cells (CHSPCs) that can be separated on the basis of AC133 expression and report that the AC133+ subset of the CHSPCs enhances the growth of tumor blood vessels in vivo in immunodeficient mice. In addition, the ratio of AC133+ proangiogenic CHSPCs to AC133 nonangiogenic CHSPCs unambiguously correlates with the severity of the clinical state of patients with peripheral arterial disease. In sum, a PFC protocol validated via in vitro and in vivo analyses, can be used to interrogate the roles of human hematopoietic elements in the growth and maintenance of the vasculature. © 2010 International Society for Advancement of Cytometry

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