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Isolation of Mouse Mammary Epithelial Subpopulations: A Comparison of Leading Methods

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Journal of Mammary Gland Biology and Neoplasia Aims and scope Submit manuscript

Abstract

Isolation of mammary epithelial subpopulations, including stem and progenitor cells, has become a standard technique in recent years. However, a number of methods and approaches for this have developed and the relative benefits of the different approaches, and the reason for their development, have not always been clear. Here, three of the leading laboratories working on the separation of mammary cell subpopulations have summarised their methods, highlighted their differences and similarities and also discussed the reasoning behind the approaches they have taken. This article will assist workers establishing mammary cell separation protocols in their laboratories to make informed choices about the methods they should use.

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Abbreviations

CFCs:

Colony Forming Cells

DAPI:

4',6-diamidino-2-phenylindole

DMEM:

Dulbecco’s Modified Eagle’s Medium

DPBS:

Dulbecco’s Phosphate Buffered Saline

EDTA:

ethylenediaminetetraacetic acid

EGTA:

ethylene glycol tetraacetic acid

F12:

Ham’s F12 Medium

FCS:

Foetal Calf Serum

FMO:

Fluorescence Minus One

FSC-H:

Forward Scatter – Height

FSC-W:

Forward Scatter – Width

HBSS:

Hank’s Balanced Salt Solution

HEPES:

hydroxyethyl piperazineethanesulfonic acid

MaSCs:

Mammary Stem Cells

MRUs:

Mammary Repopulating Units

SSC-H:

Side Scatter – Height

SSC-W:

Side Scatter – Width

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Acknowledgments

MJS and HK are supported by Cardiff University and would like to thank Kelly Soady for her assistance in protocol development. JLR is supported by Breakthrough Breast Cancer. JEV, GJL and JMS are supported by the National Health and Medical Research Council and Victorian Breast Cancer Research Consortium. JS would like to acknowledge the support of The University of Cambridge, The Breast Cancer Campaign, Hutchison Whampoa Limited and Cancer Research UK. CJW would to acknowledge the support of the Breast Cancer Campaign.

Author information

Authors and Affiliations

  1. European Cancer Stem Cell Research Institute, Cardiff School of Biosciences, Cardiff University, Museum Avenue, Cardiff, CF10 3AX, UK

    Matthew J. Smalley & Howard Kendrick

  2. Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, 3052, Australia

    Julie M. Sheridan, Geoffrey J. Lindeman & Jane E. Visvader

  3. Department of Medical Biology, University of Melbourne, Parkville, Victoria, 3010, Australia

    Julie M. Sheridan & Jane E. Visvader

  4. Breakthrough Breast Cancer Research Centre, The Institute of Cancer Research, 237 Fulham Road, London, SW3 6JB, UK

    Joseph L. Regan

  5. Cambridge Research Institute, Cancer Research UK, Li Ka Shing Centre, Robinson Way, Cambridge, CB2 0RE, UK

    Michael D. Prater & John Stingl

  6. Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QP, UK

    Christine J. Watson

  7. Department of Medicine, Royal Melbourne Hospital, University of Melbourne, Parkville, Victoria, 3010, Australia

    Geoffrey J. Lindeman

Authors
  1. Matthew J. Smalley
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  2. Howard Kendrick
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  3. Julie M. Sheridan
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  4. Joseph L. Regan
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  5. Michael D. Prater
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  6. Geoffrey J. Lindeman
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  7. Christine J. Watson
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  8. Jane E. Visvader
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  9. John Stingl
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Corresponding author

Correspondence to Matthew J. Smalley.

Additional information

Matthew J. Smalley, Jane E Visvader and John Stingl contributed equally.

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Smalley, M.J., Kendrick, H., Sheridan, J.M. et al. Isolation of Mouse Mammary Epithelial Subpopulations: A Comparison of Leading Methods. J Mammary Gland Biol Neoplasia 17, 91–97 (2012). https://doi.org/10.1007/s10911-012-9257-1

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  • DOI: https://doi.org/10.1007/s10911-012-9257-1

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