Isolation of Mouse Mammary Epithelial Subpopulations: A Comparison of Leading Methods
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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.
KeywordsMammary Stem Progenitor Epithelial Sorting Isolation
Colony Forming Cells
Dulbecco’s Modified Eagle’s Medium
Dulbecco’s Phosphate Buffered Saline
ethylene glycol tetraacetic acid
Ham’s F12 Medium
Foetal Calf Serum
Fluorescence Minus One
Forward Scatter – Height
Forward Scatter – Width
Hank’s Balanced Salt Solution
hydroxyethyl piperazineethanesulfonic acid
Mammary Stem Cells
Mammary Repopulating Units
Side Scatter – Height
Side Scatter – Width
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.
- 10.Smalley MJ. Isolation, culture and analysis of mouse mammary epithelial cells. In: Ward A, Tosh D, editors. Methods Mol Biol: Springer; 2010. p. 139–70.Google Scholar
- 11.Prater M, Shehata M, Watson CJ, Stingl J. Enzymatic dissociation, flow cytometric analysis and culture of normal mouse mammary tissue. Methods Mol Biol: Springer; In press.Google Scholar
- 17.Regan JL, Kendrick H, Magnay FA, Vafaizadeh V, Groner B, Smalley MJ. c-Kit is required for growth and survival of the cells of origin of Brca1-mutation-associated breast cancer. Oncogene. 2011. doi: 10.1038/onc.2011.289.
- 20.Shehata M, Teschendorff A, Sharp G, Novcic N, Russell IA, Avril S, et al. Phenotypic and functional characterization of the luminal cell hierarchy of the mammary gland. Submitted.Google Scholar