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

Authors
  • Galvin, Amy1
  • Weglarz, Meredith2
  • Folz-Donahue, Kat3
  • Handley, Maris1
  • Baum, Misa4
  • Mazzola, Michael5
  • Litwa, Hannah6
  • Scadden, David T5
  • Silberstein, Lev4
  • 1 HSCI-CRM Flow Cytometry Core Facility, Center for Regenerative Medicine, Massachusetts General Hospital, Boston, Massachusetts.
  • 2 Stanford Shared FACS Facility, Stanford, California.
  • 3 FACS & Imaging Core Facility, Max Planck Institute for Biology of Ageing, Cologne, Germany. , (Germany)
  • 4 Fred Hutchinson Cancer Research Center, Seattle, Washington.
  • 5 Center for Regenerative Medicine, Massachusetts General Hospital, Boston, Massachusetts.
  • 6 NYU Langone Medical Center, New York, New York.
Type
Published Article
Journal
Current protocols in cytometry
Publication Date
Jan 01, 2019
Volume
87
Issue
1
Identifiers
DOI: 10.1002/cpcy.50
PMID: 30335223
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

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. © 2018 John Wiley & Sons, Inc.

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