Cell Cycle Analysis of Hematopoietic Stem and Progenitor Cells by Multicolor Flow Cytometry

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  • Amy Galvin
    HSCI‐CRM Flow Cytometry Core Facility, Center for Regenerative Medicine, Massachusetts General Hospital Boston Massachusetts
  • Meredith Weglarz
    Stanford Shared FACS Facility Stanford California
  • Kat Folz‐Donahue
    FACS & Imaging Core Facility, Max Planck Institute for Biology of Ageing Cologne Germany
  • Maris Handley
    HSCI‐CRM Flow Cytometry Core Facility, Center for Regenerative Medicine, Massachusetts General Hospital Boston Massachusetts
  • Misa Baum
    Fred Hutchinson Cancer Research Center Seattle Washington
  • Michael Mazzola
    Center for Regenerative Medicine, Massachusetts General Hospital Boston Massachusetts
  • Hannah Litwa
    NYU Langone Medical Center New York New York
  • David T. Scadden
    Center for Regenerative Medicine, Massachusetts General Hospital Boston Massachusetts
  • Lev Silberstein
    Fred Hutchinson Cancer Research Center Seattle Washington

Abstract

<jats:title>Abstract</jats:title><jats:p>Maintenance of hematopoietic stem cell (HSC) quiescence is critical for self‐renewal and differentiation into mature lineages. Therefore, the ability to reliably detect abnormal HSC cycling is essential for experiments that seek to investigate abnormalities of HSC function. The ability to reproducibly evaluate cell cycle status in a rare cell subset requires careful optimization of multiple parameters during cell preparation and sample processing. Here, we describe a method where data acquisition parameters and fluorochrome combination for long‐term HSC staining have been specifically designed for concurrent use with DAPI and Ki‐67 antibodies. © 2018 by John Wiley & Sons, Inc.</jats:p>

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