Flow cytometry analysis of cell cycling and proliferation in mouse hematopoietic stem and progenitor cells

Barbier, Valérie, Nowlan, Bianca, Lévesque, Jean-Pierre and Winkler, Ingrid G. (2012). Flow cytometry analysis of cell cycling and proliferation in mouse hematopoietic stem and progenitor cells. In Ashman, RB (Ed.), Leucocytes: methods and protocols (pp. 31-43) New York, United States: Humana Press. doi:10.1007/978-1-61779-527-5_3


Author Barbier, Valérie
Nowlan, Bianca
Lévesque, Jean-Pierre
Winkler, Ingrid G.
Title of chapter Flow cytometry analysis of cell cycling and proliferation in mouse hematopoietic stem and progenitor cells
Title of book Leucocytes: methods and protocols
Language of Book Title eng
Place of Publication New York, United States
Publisher Humana Press
Publication Year 2012
Sub-type Chapter in textbook
DOI 10.1007/978-1-61779-527-5_3
Open Access Status DOI
Year available 2012
Series Methods in Molecular Biology
ISBN 9781617795268
9781617795275
ISSN 1064-3745
Editor Ashman, RB
Volume number 844
Chapter number 3
Start page 31
End page 43
Total pages 13
Total chapters 20
Language eng
Abstract/Summary The hematopoietic system is highly proliferative in the bone marrow (BM) due to the short half-life of granulocytes and platelets in the blood. Analysis of cell cycling and cell proliferation in vivo in specific populations of the mouse BM has highlighted some key properties of adult hematopoietic stem cells (HSCs). For instance, despite their enormous proliferation and repopulation potential, most true HSC are deeply quiescent in G(0) phase of the cell cycle and divide very infrequently, while less potent lineage-restricted progenitors divide rapidly to replace the daily consumption of blood leukocytes, erythrocytes, and platelets. In response to stress, e.g., following ablative chemotherapy or irradiation, HSC must enter the cell cycle to rapidly repopulate the BM with progenitors. Due to their extreme rarity in the BM, at least five color flow cytometry for cell surface antigens has to be combined with staining for DNA content and nuclear markers of proliferation to analyze cell cycle and proliferation of HSC in vivo. In this chapter, we describe two methods to stain mouse HSC to (1) distinguish all phases of the cell cycle (G(0), G(1), S, and G(2)/M) and (2) analyze the divisional history of HSC in vivo by incorporation of the thymidine analog 5-bromo-2-deoxyuridine.
Formatted Abstract/Summary
The hematopoietic system is highly proliferative in the bone marrow (BM) due to the short half-life of granulocytes and platelets in the blood. Analysis of cell cycling and cell proliferation in vivo in specific populations of the mouse BM has highlighted some key properties of adult hematopoietic stem cells (HSCs). For instance, despite their enormous proliferation and repopulation potential, most true HSC are deeply quiescent in G0 phase of the cell cycle and divide very infrequently, while less potent lineage-restricted progenitors divide rapidly to replace the daily consumption of blood leukocytes, erythrocytes, and platelets. In response to stress, e.g., following ablative chemotherapy or irradiation, HSC must enter the cell cycle to rapidly repopulate the BM with progenitors. Due to their extreme rarity in the BM, at least five color flow cytometry for cell surface antigens has to be combined with staining for DNA content and nuclear markers of proliferation to analyze cell cycle and proliferation of HSC in vivo. In this chapter, we describe two methods to stain mouse HSC to (1) distinguish all phases of the cell cycle (G0, G1, S, and G2/M) and (2) analyze the divisional history of HSC in vivo by incorporation of the thymidine analog 5-bromo-2-deoxyuridine.
Keyword Bone marrow
Bone marrow stroma
Flow cytometry
Hematopoietic stem cells
Q-Index Code BX
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Figures

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Created: Wed, 26 Sep 2012, 20:13:36 EST by Ms Imogen Ferrier on behalf of School of Medicine