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Use of Stem Cell Markers in Dissociated Mammary Populations

  • Dawne N. Shelton
  • Rodrigo Fernandez-Gonzalez
  • Irineu Illa-Bochaca
  • Carlos Ortiz-de-Solorzano
  • Mary Helen Barcellos-Hoff
  • Bryan E. Welm
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 621)

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.

Key words

Mammary primary culture CD24 CD49f Mammary repopulating unit 

Notes

Acknowledgments

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).

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Dawne N. Shelton
    • 1
  • Rodrigo Fernandez-Gonzalez
    • 2
  • Irineu Illa-Bochaca
    • 3
  • Carlos Ortiz-de-Solorzano
    • 4
  • Mary Helen Barcellos-Hoff
    • 3
  • Bryan E. Welm
    • 1
  1. 1.Department of SurgeryUniversity of Utah, Huntsman Cancer InstituteSalt Lake CityUSA
  2. 2.Development Biology ProgramSloan-Kettering, Institute, Memorial Sloan-Kettering Cancer CenterNew YorkUSA
  3. 3.Langone School of MedicineNew York UniversityNew YorkUSA
  4. 4.Morphology and Imaging Group and Cancer Imaging Laboratory, Center for Applied Medical ResearchUniversity of NavarrePamplonaSpain

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