Protocol

Somatic Stem Cells

Volume 879 of the series Methods in Molecular Biology pp 179-193

Date:

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

  • Abhik BandyopadhyayAffiliated withDepartment of Cellular and Structural Biology, University of Texas Health Science Center Email author 
  • , Qiaoxiang DongAffiliated withDepartment of Cellular and Structural Biology, University of Texas Health Science Center
  • , Lu-Zhe SunAffiliated withDepartment of Cellular and Structural Biology, University of Texas Health Science Center

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Abstract

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