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Use of stem cell markers in dissociated mammary populations
Dawne N Shelton1, Rodrigo Fernandez-Gonzalez, Irineu Illa-Bochaca
1Department of Surgery, University of Utah, Huntsman Cancer Institute, Salt Lake City, UT 84112, USA.
Insights
Mammary gland stem cell potential can be assessed using transplantation assays. This study details methods for isolating mammary epithelial stem cells based on specific cell surface markers for transplantation studies.
Area of Science:
- Stem cell biology
- Epithelial biology
- Mammalian development
Background:
- Mammary gland stem cells possess regenerative potential, enabling the study of epithelial stemness.
- Transplantation assays into epithelium-devoided fat pads are a key method for assessing mammary epithelial stem cell function.
- Identifying and isolating specific mammary epithelial stem cell populations is crucial for regenerative studies.
Purpose of the Study:
- To describe the materials and methods for isolating mammary epithelial stem cells using fluorescence-activated cell sorting (FACS).
- To detail the protocol for selecting cells based on markers associated with mammary repopulating potential.
- To provide a reproducible method for stem cell enrichment in mammary tissue.
Main Methods:
- Dissociation of mammary tissue into single cells.
- Staining of cells with specific cell surface markers.
- Classification and sorting of cells using fluorescence-activated cell sorting (FACS).
- Transplantation of sorted cells into epithelium-devoided recipient fat pads.
Main Results:
- The described protocol enables the isolation of mammary epithelial cell subpopulations enriched for repopulating potential.
- Successful sorting of cells based on established and novel cell surface markers is demonstrated.
- The methodology provides a foundation for comparative studies of mammary stem cell populations.
Conclusions:
- The described FACS-based method is effective for isolating mammary epithelial stem cells with regenerative potential.
- This protocol can be adapted to utilize various cell surface markers for stem cell enrichment.
- The study provides a valuable resource for researchers investigating mammary gland development and regeneration.
Abstract:
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.
