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.
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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