Stem/Progenitor Cells in Murine Mammary Gland: Isolation and Functional Characterization

Part of the Methods in Molecular Biology book series (MIMB, volume 879)


The presence of adult functional mammary epithelial stem/progenitor cells in mammary gland and recent identification of enriched fraction of transplantable mammary stem cells (MaSCs) through specific cell surface markers have revolutionized the study to delineate the role of mammary stem/progenitor functions in both mammary gland development and mammary tumorigenesis. In this chapter, we have described detailed methods of isolation of mammary epithelial cells from murine mammary glands, enrichment of stem/progenitor fractions by fluorescence-activated cell sorting (FACS), in vitro mammosphere culture, and differentiation assays in two- and three-dimensional culture. We have also described the detailed protocol of in vivo cleared mammary fat pad transplantation assay for the assessment of the MaSC repopulating activity, which indicates their multilineage differentiation and self-renewal potential in vivo and is considered the “gold standard” assay of functional stem cells.

Key words

Mammary stem cells Mammary progenitor cells Mammary glands Cleared mammary fat pad Mammosphere Mice Fluorescence-activated cell sorting 



This work was supported in part by NIH grants R01CA079683, R01CA075253 (L-ZS), and Shelby Rae Tangg foundation (AB). We thank the Flow Cytometry Shared Resource Facility and department of Pathology of UT Health Science Center at San Antonio for their assistance in analysis and interpretation of the results.


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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Abhik Bandyopadhyay
    • 1
  • Qiaoxiang Dong
    • 1
  • Lu-Zhe Sun
    • 1
  1. 1.Department of Cellular and Structural BiologyUniversity of Texas Health Science CenterSan AntonioUSA

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