Get access

Flow cytometric techniques for detection of candidate cancer stem cell subpopulations in canine tumour models

Authors

  • T. M. Blacking,

    Corresponding author
    1. Comparative Oncology and Stem Cell Research Group, The Royal (Dick) School of Veterinary Studies and Roslin Institute, The University of Edinburgh, Easter Bush, Midlothian, Edinburgh, UK
    Search for more papers by this author
  • M. Waterfall,

    1. Flow Cytometry Facility, Ashworth Laboratories, Institute of Immunology and Infection Research, The University of Edinburgh, School of Biological Sciences, Kings' Buildings, Edinburgh, UK
    Search for more papers by this author
  • K. Samuel,

    1. MRC Centre for Regenerative Medicine, The University of Edinburgh, Edinburgh, UK
    Search for more papers by this author
  • D. J. Argyle

    1. Comparative Oncology and Stem Cell Research Group, The Royal (Dick) School of Veterinary Studies and Roslin Institute, The University of Edinburgh, Easter Bush, Midlothian, Edinburgh, UK
    Search for more papers by this author

T. M. Blacking
The Royal (Dick) School of Veterinary Studies and Roslin Institute
The University of Edinburgh
Easter Bush, Midlothian
Edinburgh EH25 9RG, UK
e-mail: thaliablacking@hotmail.com

Abstract

The cancer stem cell (CSC) hypothesis proposes that tumour growth is maintained by a distinct subpopulation of ‘CSC’. This study applied flow cytometric methods, reported to detect CSC in both primary and cultured cancer cells of other species, to identify candidate canine subpopulations. Cell lines representing diverse canine malignancies, and cells derived from spontaneous canine tumours, were evaluated for expression of stem cell-associated surface markers (CD34, CD44, CD117 and CD133) and functional properties [Hoecsht 33342 efflux, aldehyde dehydrogenase (ALDH) activity]. No discrete marker-defined subsets were identified within established cell lines; cells derived directly from spontaneous tumours demonstrated more heterogeneity, although this diminished upon in vitro culture. Functional assays produced variable results, suggesting context-dependency. Flow cytometric methods may be adopted to identify putative canine CSC. Whilst cell lines are valuable in assay development, primary cells may provide a more rewarding model for studying tumour heterogeneity in the context of CSC. However, it will be essential to fully characterize any candidate subpopulations to ensure that they meet CSC criteria.

Ancillary