Distinct stem cells contribute to mammary gland development and maintenance


The mammary epithelium is composed of several cell lineages including luminal, alveolar and myoepithelial cells. Transplantation studies have suggested that the mammary epithelium is maintained by the presence of multipotent mammary stem cells. To define the cellular hierarchy of the mammary gland during physiological conditions, we performed genetic lineage-tracing experiments and clonal analysis of the mouse mammary gland during development, adulthood and pregnancy. We found that in postnatal unperturbed mammary gland, both luminal and myoepithelial lineages contain long-lived unipotent stem cells that display extensive renewing capacities, as demonstrated by their ability to clonally expand during morphogenesis and adult life as well as undergo massive expansion during several cycles of pregnancy. The demonstration that the mammary gland contains different types of long-lived stem cells has profound implications for our understanding of mammary gland physiology and will be instrumental in unravelling the cells at the origin of breast cancers.

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Figure 1: All mammary epithelial lineages derive from embryonic K14-expressing progenitors.
Figure 2: K14-expressing stem cells ensure mammary myoepithelial lineage expansion and maintenance during puberty, adult life and pregnancy.
Figure 3: K8-expressing stem cells ensure mammary luminal lineage expansion and maintenance during puberty, adult life and pregnancy.
Figure 4: Myoepithelial and luminal stem cells maintain their unipotent fate in mammary reconstitution assay when co-transplanted in non-limiting conditions.
Figure 5: Myoepithelial but not luminal stem cells can be forced to adopt a multipotent fate in mammary reconstitution assays.


  1. 1

    Watson, C. J. & Khaled, W. T. Mammary development in the embryo and adult: a journey of morphogenesis and commitment. Development 135, 995–1003 (2008)

    CAS  Article  Google Scholar 

  2. 2

    Smalley, M. & Ashworth, A. Stem cells and breast cancer: A field in transit. Nature Rev. Cancer 3, 832–844 (2003)

    CAS  Article  Google Scholar 

  3. 3

    Visvader, J. E. Keeping abreast of the mammary epithelial hierarchy and breast tumorigenesis. Genes Dev. 23, 2563–2577 (2009)

    CAS  Article  Google Scholar 

  4. 4

    Visvader, J. E. & Smith, G. H. Murine mammary epithelial stem cells: Discovery, function, and current status. Cold Spring Harb. Perspect. Biol. 3, 1–14 (2011)

    Article  Google Scholar 

  5. 5

    Stingl, J. Detection and analysis of mammary gland stem cells. J. Pathol. 217, 229–241 (2009)

    CAS  Article  Google Scholar 

  6. 6

    Pechoux, C., Gudjonsson, T., Ronnov-Jessen, L., Bissell, M. J. & Petersen, O. W. Human mammary luminal epithelial cells contain progenitors to myoepithelial cells. Dev. Biol. 206, 88–99 (1999)

    CAS  Article  Google Scholar 

  7. 7

    Stingl, J., Eaves, C. J., Kuusk, U. & Emerman, J. T. Phenotypic and functional characterization in vitro of a multipotent epithelial cell present in the normal adult human breast. Differentiation 63, 201–213 (1998)

    CAS  Article  Google Scholar 

  8. 8

    Gudjonsson, T. et al. Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties. Genes Dev. 16, 693–706 (2002)

    CAS  Article  Google Scholar 

  9. 9

    Dontu, G. et al. In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. Genes Dev. 17, 1253–1270 (2003)

    CAS  Article  Google Scholar 

  10. 10

    Kordon, E. C. & Smith, G. H. An entire functional mammary gland may comprise the progeny from a single cell. Development 125, 1921–1930 (1998)

    CAS  PubMed  Google Scholar 

  11. 11

    Smith, G. H. Experimental mammary epithelial morphogenesis in an in vivo model: evidence for distinct cellular progenitors of the ductal and lobular phenotype. Breast Cancer Res. Treat. 39, 21–31 (1996)

    CAS  Article  Google Scholar 

  12. 12

    Stingl, J. et al. Purification and unique properties of mammary epithelial stem cells. Nature 439, 993–997 (2006)

    CAS  Article  ADS  Google Scholar 

  13. 13

    Shackleton, M. et al. Generation of a functional mammary gland from a single stem cell. Nature 439, 84–88 (2006)

    CAS  Article  ADS  Google Scholar 

  14. 14

    Blanpain, C. & Fuchs, E. Epidermal homeostasis: a balancing act of stem cells in the skin. Nature Rev. Mol. Cell Biol. 10, 207–217 (2009)

    CAS  Article  Google Scholar 

  15. 15

    Sleeman, K. E., Kendrick, H., Ashworth, A., Isacke, C. M. & Smalley, M. J. CD24 staining of mouse mammary gland cells defines luminal epithelial, myoepithelial/basal and non-epithelial cells. Breast Cancer Res. 8, R7 (2006)

    Article  Google Scholar 

  16. 16

    Lim, E. et al. Transcriptome analyses of mouse and human mammary cell subpopulations reveal multiple conserved genes and pathways. Breast Cancer Res. 12, R21 (2010)

    Article  Google Scholar 

  17. 17

    Kendrick, H. et al. Transcriptome analysis of mammary epithelial subpopulations identifies novel determinants of lineage commitment and cell fate. BMC Genomics 9, 591 (2008)

    Article  Google Scholar 

  18. 18

    Barker, N. et al. Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature 449, 1003–1007 (2007)

    CAS  Article  ADS  Google Scholar 

  19. 19

    Wagner, K. U. et al. An adjunct mammary epithelial cell population in parous females: its role in functional adaptation and tissue renewal. Development 129, 1377–1386 (2002)

    CAS  PubMed  Google Scholar 

  20. 20

    Booth, B. W., Boulanger, C. A. & Smith, G. H. Alveolar progenitor cells develop in mouse mammary glands independent of pregnancy and lactation. J. Cell. Physiol. 212, 729–736 (2007)

    CAS  Article  Google Scholar 

  21. 21

    Chepko, G. & Smith, G. H. Three division-competent, structurally-distinct cell populations contribute to murine mammary epithelial renewal. Tissue Cell 29, 239–253 (1997)

    CAS  Article  Google Scholar 

  22. 22

    Rudland, P. S. & Hughes, C. M. Immunocytochemical identification of cell types in human mammary gland: variations in cellular markers are dependent on glandular topography and differentiation. J. Histochem. Cytochem. 37, 1087–1100 (1989)

    CAS  Article  Google Scholar 

  23. 23

    Fernandez-Gonzalez, R. et al. Mapping mammary gland architecture using multi-scale in situ analysis. Integr. Biol. 1, 80–89 (2009)

    CAS  Article  Google Scholar 

  24. 24

    Van Keymeulen, A. et al. Epidermal progenitors give rise to Merkel cells during embryonic development and adult homeostasis. J. Cell Biol. 187, 91–100 (2009)

    CAS  Article  Google Scholar 

  25. 25

    Liu, X. et al. Somatic loss of BRCA1 and p53 in mice induces mammary tumors with features of human BRCA1-mutated basal-like breast cancer. Proc. Natl Acad. Sci. USA 104, 12111–12116 (2007)

    CAS  Article  ADS  Google Scholar 

  26. 26

    Srinivasan, K., Strickland, P., Valdes, A., Shin, G. C. & Hinck, L. Netrin-1/neogenin interaction stabilizes multipotent progenitor cap cells during mammary gland morphogenesis. Dev. Cell 4, 371–382 (2003)

    CAS  Article  Google Scholar 

  27. 27

    Naylor, S. et al. Retroviral expression of Wnt-1 and Wnt-7b produces different effects in mouse mammary epithelium. J. Cell Sci. 113, 2129–2138 (2000)

    CAS  PubMed  Google Scholar 

  28. 28

    Welm, B. E. et al. Sca-1pos cells in the mouse mammary gland represent an enriched progenitor cell population. Dev. Biol. 245, 42–56 (2002)

    CAS  Article  Google Scholar 

  29. 29

    Welm, B. E., Dijkgraaf, G. J., Bledau, A. S., Welm, A. L. & Werb, Z. Lentiviral transduction of mammary stem cells for analysis of gene function during development and cancer. Cell Stem Cell 2, 90–102 (2008)

    CAS  Article  Google Scholar 

  30. 30

    Tsai, Y. C. et al. Contiguous patches of normal human mammary epithelium derived from a single stem cell: implications for breast carcinogenesis. Cancer Res. 56, 402–404 (1996)

    CAS  PubMed  Google Scholar 

  31. 31

    Badders, N. M. et al. The Wnt receptor, Lrp5, is expressed by mouse mammary stem cells and is required to maintain the basal lineage. PLoS ONE 4, e6594 (2009)

    Article  ADS  Google Scholar 

  32. 32

    Taddei, I. et al. β1 integrin deletion from the basal compartment of the mammary epithelium affects stem cells. Nature Cell Biol. 10, 716–722 (2008)

    CAS  Article  Google Scholar 

  33. 33

    Srinivas, S. et al. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev. Biol. 1, 4 (2001)

    CAS  Article  Google Scholar 

  34. 34

    Madisen, L. et al. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nature Neurosci. 13, 133–140 (2010)

    CAS  Article  Google Scholar 

  35. 35

    Vasioukhin, V., Bauer, C., Degenstein, L., Wise, B. & Fuchs, E. Hyperproliferation and defects in epithelial polarity upon conditional ablation of α-catenin in skin. Cell 104, 605–617 (2001)

    CAS  Article  Google Scholar 

  36. 36

    Nguyen, H., Rendl, M. & Fuchs, E. Tcf3 governs stem cell features and represses cell fate determination in skin. Cell 127, 171–183 (2006)

    CAS  Article  Google Scholar 

  37. 37

    Perl, A. K., Wert, S. E., Nagy, A., Lobe, C. G. & Whitsett, J. A. Early restriction of peripheral and proximal cell lineages during formation of the lung. Proc. Natl Acad. Sci. USA 99, 10482–10487 (2002)

    CAS  Article  ADS  Google Scholar 

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We thank our colleagues who provided us with reagents, which are cited in the text, and B. Hogan for sharing unpublished mice. We thank our colleagues from the Blanpain laboratory and C. Govaerts for their comments on the manuscript. We thank J. Rosen for discussion and M. Van Lohuizen and K. Nacerddine for their help with the transplantation assay. We thank F. Bollet-Quivogne and J.-M. Vanderwinden for their help with confocal imaging. C.B. and A.V.K. are chercheur qualifié, B.B. is chargé de recherche and M.O. is a collaborateur scientifique of the FRS/FNRS. A.S.R. is supported by TELEVIE and the Portuguese Science Foundation (FCT). N.S. is supported by the Fondation Contre le Cancer. J.R. is supported by the grant F32HL102920. C.B. is an investigator of Welbio. This work was supported by the FNRS, TELEVIE, the program d’excellence CIBLES of the Wallonia Region, a research grant from the Fondation Contre le Cancer, the ULB fondation, the fond Gaston Ithier, a starting grant of the European Research Council (ERC) and the EMBO Young Investigator Program.

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C.B., A.V.K, A.S.R. designed the experiments and performed data analysis. AV.K., A.S.R. and M.O. performed most of the experiments, J.R. generated the K5-CreER knockin mice, B.B., S.D. and A.V.K. performed the FACS analysis and cell sorting. G.B. and N.S. provided technical support. C.B. wrote the manuscript.

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Correspondence to Cédric Blanpain.

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The authors declare no competing financial interests.

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Van Keymeulen, A., Rocha, A., Ousset, M. et al. Distinct stem cells contribute to mammary gland development and maintenance. Nature 479, 189–193 (2011). https://doi.org/10.1038/nature10573

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