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

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Somatic Stem Cells

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

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.

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Acknowledgments

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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Authors and Affiliations

  1. Department of Cellular and Structural Biology, University of Texas Health Science Center, San Antonio, TX, USA

    Abhik Bandyopadhyay, Qiaoxiang Dong & Lu-Zhe Sun

Authors
  1. Abhik Bandyopadhyay
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  2. Qiaoxiang Dong
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  3. Lu-Zhe Sun
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Corresponding author

Correspondence to Abhik Bandyopadhyay .

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Editors and Affiliations

  1. Mouse Cancer Genetics Program, National Cancer Institute, Center Drive 31, Frederick, 21702, Maryland, USA

    Shree Ram Singh

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© 2012 Springer Science+Business Media, LLC

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Bandyopadhyay, A., Dong, Q., Sun, LZ. (2012). Stem/Progenitor Cells in Murine Mammary Gland: Isolation and Functional Characterization. In: Singh, S. (eds) Somatic Stem Cells. Methods in Molecular Biology, vol 879. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-815-3_12

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  • DOI: https://doi.org/10.1007/978-1-61779-815-3_12

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-814-6

  • Online ISBN: 978-1-61779-815-3

  • eBook Packages: Springer Protocols

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