Use of Stem Cell Markers in Dissociated Mammary Populations
The regenerative potential of mammary epithelium facilitates assessment of the “stemness” of any epithelial subpopulation in transplantation assays. Thus, mammary tissue can be dissociated into single cells, stained for cell surface markers of interest and classified using fluorescence-activated cell sorting. The selected cells can then be transplanted into epithelium-devoided fat pads from recipient hosts. Recent publications have described markers that enrich for mammary repopulating potential. Here, we describe the materials and methods necessary to sort cells according to these markers. This approach can be used interchangeably with other cell surface markers with slight variation to the protocol.
Key wordsMammary primary culture CD24 CD49f Mammary repopulating unit
This work was supported by a predoctoral fellowship to RFG from the Department of Defense Breast Cancer Research Program (DAMD 17-03-1-0594), grants from the same institution to COS (DAMD 17-00-1-0227 and DAMD 17-00-1-0306), a grant to BEW from the National Cancer Institute (CA 8424306), and a grant to MHBH funded by the National Institute of Environmental Health Sciences and the National Cancer Institute (U01ES012801).
- 6.Asselin-Labat ML, Sutherland KD, Barker H, Thomas R, Shackleton M, Forrest NC, Hartley L, Robb L, Grosveld FG, van der Wees J, Lindeman GJ, Visvader JE (2007) Gata-3 is an essential regulator of mammary-gland morphogenesis and luminal-cell differentiation. Nat Cell Biol 9:201–209CrossRefPubMedGoogle Scholar