Stem Cell Reviews and Reports

, Volume 9, Issue 5, pp 731–739

Optimised Protocols for the Identification of the Murine Cardiac Side Population

  • Annette Meeson
  • Andrew Fuller
  • David T. Breault
  • W. Andrew Owens
  • Gavin D. Richardson
Article

DOI: 10.1007/s12015-013-9440-9

Cite this article as:
Meeson, A., Fuller, A., Breault, D.T. et al. Stem Cell Rev and Rep (2013) 9: 731. doi:10.1007/s12015-013-9440-9
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Abstract

Cardiac side population (CSP) cells, defined by their ability to efflux the vital dye Hoechst 33342, have been identified as putative cardiac stem cells based on their potential to give rise to both cardiomyocytes and endothelial cells. The CSP phenotype relies on an active metabolic pathway and cell viability to identify a rare population of cells and therefore technical differences in the CSP staining protocol can lead to inconsistent results and discrepancies between studies. Here we describe an established protocol for CSP identification and have optimised a protocol for CSP analysis utilizing an automated cardiac digestion procedure using gentleMACs dissociation and Hoechst 33342 staining followed by dual wavelength flow cytometric analysis.

Keywords

Cardiac Side population Abcg2 Fumitremorgin C Stem cell 

Abbreviations

FACS

Fluorescence-activated cell sorting

SP

Side population

CSP

Cardiac side population

FTC

Fumitremorgin C

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Annette Meeson
    • 1
  • Andrew Fuller
    • 1
  • David T. Breault
    • 3
    • 4
  • W. Andrew Owens
    • 1
    • 2
  • Gavin D. Richardson
    • 1
  1. 1.Institute of Genetic Medicine, International Centre for LifeNewcastle UniversityNewcastle upon TyneUK
  2. 2.Department of Cardiothoracic SurgerySouth Tees Hospitals NHS Foundation TrustMiddlesbroughUK
  3. 3.Division of Endocrinology, Children’s Hospital BostonHarvard Medical SchoolBostonUSA
  4. 4.Harvard Stem Cell InstituteCambridgeUSA