Mammary Epithelial Stem Cells

  • Brian W. Booth
  • Daniel Medina
  • Gilbert H. Smith


An entire mammary epithelial outgrowth, capable of full secretory differentiation, may comprise the progeny of a single stem cell. Experimental evidence has indicated three distinct mammary progenitor subtypes: unlimited progenitor cells, duct-limited progenitors, and lobule-limited progenitor cells. These different progenitor cell types drive the individual stages of mammary development taking direction from the surrounding microenvironment and systemic hormones. Mammary epithelial stem cells reside in basal positions, in contact with the basement membrane, but not in luminal positions throughout the mammary ductal tree. Stem cell morphology is characterized by the absence of cellular organelles resulting in their light or pale appearance in thin and ultrathin tissue sections. In recent years mammary stem and progenitor cells have been isolated based on the expression of cell surface markers, thus allowing increased study of these cells. The purpose of this chapter is to summarize the experimental history of mammary stem cells and to address future areas and challenges for the investigation of these cells.


Mammary Gland Mammary Epithelial Cell Mammary Epithelium Human Mammary Epithelial Cell Mammary Stem 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.





Differentiating large light cells


Estrogen receptor


Mouse mammary tumor virus promoter


Mammary repopulating unit


Parity-identified mammary epithelial cells


Progesterone receptor


Small light cells


Terminal ductal lobule unit


Undifferentiated large light cells


Whey acidic protein promoter



Figure 3 was illustrated by Eve E. Kingsley Booth.


  1. 1.
    DeOme KB, Faulkin Jr LJ, Bern HA, Blair PB. Development of mammary tumors from hyperplastic alveolar nodules transplanted into gland-free mammary fat pads of female C3H mice. Cancer Res. 1959;19:515–20.PubMedGoogle Scholar
  2. 2.
    Faulkin Jr LJ, Deome KB. Regulation of growth and spacing of gland elements in the mammary fat pad of the C3H mouse. J Natl Cancer Inst. 1960;24:953–69.PubMedGoogle Scholar
  3. 3.
    Daniel CW, Aidells BD, Medina D, Faulkin Jr LJ. Unlimited division potential of precancerous mouse mammary cells after spontaneous or carcinogen-induced transformation. Fed Proc. 1975;34:64–7.PubMedGoogle Scholar
  4. 4.
    Callahan R, Smith GH. MMTV-induced mammary tumorigenesis: gene discovery, progression to malignancy and cellular pathways. Oncogene. 2000;19:992–1001.PubMedCrossRefGoogle Scholar
  5. 5.
    Smith GH, Pauley RJ, Socher SH, Medina D. Chemical carcinogenesis in C3H/StWi mice, a worthwhile experimental model for breast cancer. Cancer Res. 1978;38:4504–9.PubMedGoogle Scholar
  6. 6.
    Smith GH, Arthur LA, Medina D. Evidence of separate pathways for viral and chemical carcinogenesis in C3H/StWi mouse mammary glands. Int J Cancer. 1980;26:373–9.PubMedCrossRefGoogle Scholar
  7. 7.
    Medina D. The preneoplastic phenotype in murine mammary tumorigenesis. J Mammary Gland Biol Neoplasia. 2000;5: 393–407.PubMedCrossRefGoogle Scholar
  8. 8.
    Medina D. Biological and molecular characteristics of the premalignant mouse mammary gland. Biochim Biophys Acta. 2002;1603:1–9.PubMedGoogle Scholar
  9. 9.
    Maglione JE, Moghanaki D, Young LJ, et al. Transgenic Polyoma middle-T mice model premalignant mammary disease. Cancer Res. 2001;61:8298–305.PubMedGoogle Scholar
  10. 10.
    Medina D, Kittrell FS, Shepard A, et al. Biological and genetic properties of the p53 null preneoplastic mammary epithelium. FASEB J. 2002;16:881–3.PubMedGoogle Scholar
  11. 11.
    Medina D, Kittrell FS. Immortalization phenotype dissociated from the preneoplastic phenotype in mouse mammary epithelial outgrowths in vivo. Carcinogenesis. 1993;14:25–8.PubMedCrossRefGoogle Scholar
  12. 12.
    Medina D, Kittrell FS, Liu YJ, Schwartz M. Morphological and functional properties of TM preneoplastic mammary outgrowths. Cancer Res. 1993;53:663–7.PubMedGoogle Scholar
  13. 13.
    Young LJ, Medina D, DeOme KB, Daniel CW. The influence of host and tissue age on life span and growth rate of serially transplanted mouse mammary gland. Exp Gerontol. 1971;6:49–56.PubMedCrossRefGoogle Scholar
  14. 14.
    Smith GH, Medina D. A morphologically distinct candidate for an epithelial stem cell in mouse mammary gland. J Cell Sci. 1988;90(Pt 1):173–83.PubMedGoogle Scholar
  15. 15.
    Smith GH. Experimental mammary epithelial morphogenesis in an in vivo model: evidence for distinct cellular progenitors of the ductal and lobular phenotype. Breast Cancer Res Treat. 1996;39: 21–31.PubMedCrossRefGoogle Scholar
  16. 16.
    Daniel CW, Young LJ. Influence of cell division on an aging process. Life span of mouse mammary epithelium during serial propagation in vivo. Exp Cell Res. 1971;65:27–32.PubMedCrossRefGoogle Scholar
  17. 17.
    Chepko G, Smith GH. Three division-competent, structurally-distinct cell populations contribute to murine mammary epithelial renewal. Tissue Cell. 1997;29:239–53.PubMedCrossRefGoogle Scholar
  18. 18.
    Smith GH, Vonderhaar BK. Functional differentiation in mouse mammary gland epithelium is attained through DNA synthesis, inconsequent of mitosis. Dev Biol. 1981;88:167–79.PubMedCrossRefGoogle Scholar
  19. 19.
    Vonderhaar BK, Smith GH. Dissociation of cytological and functional differential in virgin mouse mammary gland during inhibition of DNA synthesis. J Cell Sci. 1982;53:97–114.PubMedGoogle Scholar
  20. 20.
    Graham DE, Medina D, Smith GH. Increased concentration of an indigenous proviral mouse mammary tumor virus long terminal repeat-containing transcript is associated with neoplastic transformation of mammary epithelium in C3H/Sm mice. J Virol. 1984;49: 819–27.PubMedGoogle Scholar
  21. 21.
    Nicoll CS, Tucker HA. Estimates of parenchymal, stromal, and lymph node deoxyribonucleic acid in mammary glands of C3H/Crgl-2 mice. Life Sci. 1965;4:993–1001.PubMedCrossRefGoogle Scholar
  22. 22.
    Kordon EC, Smith GH. An entire functional mammary gland may comprise the progeny from a single cell. Development. 1998;125:1921–30.PubMedGoogle Scholar
  23. 23.
    Smith GH, Strickland P, Daniel CW. Putative epithelial stem cell loss corresponds with mammary growth senescence. Cell Tissue Res. 2002;310:313–20.PubMedCrossRefGoogle Scholar
  24. 24.
    Gudjonsson T, Villadsen R, Nielsen HL, Ronnov-Jessen L, Bissell MJ, Petersen OW. Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties. Genes Dev. 2002;16:693–706.PubMedCrossRefGoogle Scholar
  25. 25.
    Smith GH, Chepko G. Mammary epithelial stem cells. Microsc Res Tech. 2001;52:190–203.PubMedCrossRefGoogle Scholar
  26. 26.
    Kim ND, Clifton KH. Characterization of rat mammary epithelial cell subpopulations by peanut lectin and anti-Thy-1.1 antibody and study of flow-sorted cells in vivo. Exp Cell Res. 1993;207:74–85.PubMedCrossRefGoogle Scholar
  27. 27.
    Kamiya K, Gould MN, Clifton KH. Quantitative studies of ductal versus alveolar differentiation from rat mammary clonogens. Proc Soc Exp Biol Med. 1998;219:217–25.PubMedCrossRefGoogle Scholar
  28. 28.
    Smith GH, Boulanger CA. Mammary stem cell repertoire: new insights in aging epithelial populations. Mech Ageing Dev. 2002;123:1505–19.PubMedCrossRefGoogle Scholar
  29. 29.
    Wagner KU, Boulanger CA, Henry MD, Sgagias M, Hennighausen L, Smith GH. An adjunct mammary epithelial cell population in parous females: its role in functional adaptation and tissue renewal. Development. 2002;129:1377–86.PubMedGoogle Scholar
  30. 30.
    Boulanger CA, Wagner KU, Smith GH. Parity-induced mouse mammary epithelial cells are pluripotent, self-renewing and sensitive to TGF-beta1 expression. Oncogene. 2005;24:552–60.PubMedCrossRefGoogle Scholar
  31. 31.
    Booth BW, Boulanger CA, Smith GH. Alveolar progenitor cells develop in mouse mammary glands independent of pregnancy and lactation. J Cell Physiol. 2007;212:729–36.PubMedCrossRefGoogle Scholar
  32. 32.
    Matulka LA, Triplett AA, Wagner KU. Parity-induced mammary epithelial cells are multipotent and express cell surface markers associated with stem cells. Dev Biol. 2007;303:29–44.PubMedCrossRefGoogle Scholar
  33. 33.
    Stingl J, Eirew P, Ricketson I, et al. Purification and unique properties of mammary epithelial stem cells. Nature. 2006;439:993–7.PubMedGoogle Scholar
  34. 34.
    Gusterson BA, Williams J, Bunnage H, O’Hare MJ, Dubois JD. Human breast epithelium transplanted into nude mice. Proliferation and milk protein production in response to pregnancy. Virchows Arch A Pathol Anat Histopathol. 1984;404:325–33.PubMedCrossRefGoogle Scholar
  35. 35.
    Knight J, Gusterson BA, Cowley G, Monaghan P. Differentiation of normal and malignant human squamous epithelium in vivo and in vitro: a morphologic study. Ultrastruct Pathol. 1984;7:133–41.PubMedCrossRefGoogle Scholar
  36. 36.
    Sheffield LG, Welsch CW. Transplantation of human breast epithelia to mammary-gland-free fat-pads of athymic nude mice: influence of mammotrophic hormones on growth of breast epithelia. Int J Cancer. 1988;41:713–9.PubMedCrossRefGoogle Scholar
  37. 37.
    Hovey RC, McFadden TB, Akers RM. Regulation of mammary gland growth and morphogenesis by the mammary fat pad: a species comparison. J Mammary Gland Biol Neoplasia. 1999;4: 53–68.PubMedCrossRefGoogle Scholar
  38. 38.
    Kuperwasser C, Chavarria T, Wu M, et al. Reconstruction of functionally normal and malignant human breast tissues in mice. Proc Natl Acad Sci U S A. 2004;101:4966–71.PubMedCrossRefGoogle Scholar
  39. 39.
    Proia DA, Kuperwasser C. Reconstruction of human mammary tissues in a mouse model. Nat Protoc. 2006;1:206–14.PubMedCrossRefGoogle Scholar
  40. 40.
    Eirew P, Stingl J, Raouf A, et al. A method for quantifying normal human mammary epithelial stem cells with in vivo regenerative ability. Nat Med. 2008;14:1384–9.PubMedCrossRefGoogle Scholar
  41. 41.
    Shackleton M, Vaillant F, Simpson KJ, et al. Generation of a functional mammary gland from a single stem cell. Nature. 2006;439:84–8.PubMedCrossRefGoogle Scholar
  42. 42.
    Taddei I, Deugnier MA, Faraldo MM, et al. Beta1 integrin deletion from the basal compartment of the mammary epithelium affects stem cells. Nat Cell Biol. 2008;10:716–22.PubMedCrossRefGoogle Scholar
  43. 43.
    Van Keymeulen A, Rocha AS, Ousset M, et al. Distinct stem cells contribute to mammary gland development and maintenance. Nature. 2011;479:189–93.PubMedCrossRefGoogle Scholar
  44. 44.
    Gupta PB, Fillmore CM, Jiang G, et al. Stochastic state transitions give rise to phenotypic equilibrium in populations of cancer cells. Cell. 2011;146:633–44.PubMedCrossRefGoogle Scholar
  45. 45.
    Sleeman KE, Kendrick H, Ashworth A, Isacke CM, Smalley MJ. CD24 staining of mouse mammary gland cells defines luminal epithelial, myoepithelial/basal and non-epithelial cells. Breast Cancer Res. 2006;8:R7.PubMedCrossRefGoogle Scholar
  46. 46.
    Cremers N, Deugnier MA, Sleeman J. Loss of CD24 expression promotes ductal branching in the murine mammary gland. Cell Mol Life Sci. 2010;67:2311–22.PubMedCrossRefGoogle Scholar
  47. 47.
    Villadsen R, Fridriksdottir AJ, Ronnov-Jessen L, et al. Evidence for a stem cell hierarchy in the adult human breast. J Cell Biol. 2007;177:87–101.PubMedCrossRefGoogle Scholar
  48. 48.
    Stingl J, Eaves CJ, Kuusk U, Emerman JT. Phenotypic and functional characterization in vitro of a multipotent epithelial cell present in the normal adult human breast. Differentiation. 1998;63:201–13.PubMedCrossRefGoogle Scholar
  49. 49.
    Stingl J, Eaves CJ, Zandieh I, Emerman JT. Characterization of bipotent mammary epithelial progenitor cells in normal adult human breast tissue. Breast Cancer Res Treat. 2001;67:93–109.PubMedCrossRefGoogle Scholar
  50. 50.
    Raouf A, Zhao Y, To K, et al. Transcriptome analysis of the normal human mammary cell commitment and differentiation process. Cell Stem Cell. 2008;3:109–18.PubMedCrossRefGoogle Scholar
  51. 51.
    Carter WG, Kaur P, Gil SG, Gahr PJ, Wayner EA. Distinct functions for integrins alpha 3 beta 1 in focal adhesions and alpha 6 beta 4/bullous pemphigoid antigen in a new stable anchoring contact (SAC) of keratinocytes: relation to hemidesmosomes. J Cell Biol. 1990;111:3141–54.PubMedCrossRefGoogle Scholar
  52. 52.
    Moraes RC, Zhang X, Harrington N, et al. Constitutive activation of smoothened (SMO) in mammary glands of transgenic mice leads to increased proliferation, altered differentiation and ductal dysplasia. Development. 2007;134:1231–42.PubMedCrossRefGoogle Scholar
  53. 53.
    Vaillant F, Lindeman GJ, Visvader JE. Jekyll or Hyde: does Matrigel provide a more or less physiological environment in mammary repopulating assays? Breast Cancer Res. 2011;13:108.PubMedCrossRefGoogle Scholar
  54. 54.
    Chew EC, Hoshino K. Early histogenesis of transplanted mouse mammary glands. II. Within 96 hours following isografting. Z Anat Entwicklungsgesch. 1970;132:318–24.PubMedCrossRefGoogle Scholar
  55. 55.
    Medina D, Vaage J, Sedlacek R. Mammary noduligenesis and tumorigenesis in pathogen-free C3Hf mice. J Natl Cancer Inst. 1973;51:961–5.PubMedGoogle Scholar
  56. 56.
    Mallepell S, Krust A, Chambon P, Brisken C. Paracrine signaling through the epithelial estrogen receptor alpha is required for proliferation and morphogenesis in the mammary gland. Proc Natl Acad Sci U S A. 2006;103:2196–201.PubMedCrossRefGoogle Scholar
  57. 57.
    Ciarloni L, Mallepell S, Brisken C. Amphiregulin is an essential mediator of estrogen receptor alpha function in mammary gland development. Proc Natl Acad Sci U S A. 2007;104:5455–60.PubMedCrossRefGoogle Scholar
  58. 58.
    Brisken C, Park S, Vass T, Lydon JP, O’Malley BW, Weinberg RA. A paracrine role for the epithelial progesterone receptor in mammary gland development. Proc Natl Acad Sci U S A. 1998;95: 5076–81.PubMedCrossRefGoogle Scholar
  59. 59.
    Raafat A, Strizzi L, Lashin K, et al. Effects of age and parity on mammary gland lesions and progenitor cells in the FVB/N-RC mice. PLoS One. 2012;7:e43624.PubMedCrossRefGoogle Scholar
  60. 60.
    Daniel CW, Young LJ, Medina D, DeOme KB. The influence of mammogenic hormones on serially transplanted mouse mammary gland. Exp Gerontol. 1971;6:95–101.PubMedCrossRefGoogle Scholar
  61. 61.
    Boulanger CA, Mack DL, Booth BW, Smith GH. Interaction with the mammary microenvironment redirects spermatogenic cell fate in vivo. Proc Natl Acad Sci U S A. 2007;104:3871–6.PubMedCrossRefGoogle Scholar
  62. 62.
    Booth BW, Mack DL, Androutsellis-Theotokis A, McKay RD, Boulanger CA, Smith GH. The mammary microenvironment alters the differentiation repertoire of neural stem cells. Proc Natl Acad Sci U S A. 2008;105:14891–6.PubMedCrossRefGoogle Scholar
  63. 63.
    Boulanger CA, Bruno RD, Rosu-Myles M, Smith GH. The mouse mammary microenvironment redirects mesoderm-derived bone marrow cells to a mammary epithelial progenitor cell fate. Stem Cells Dev. 2011;21:948–54.PubMedCrossRefGoogle Scholar
  64. 64.
    Bussard KM, Boulanger CA, Booth BW, Bruno RD, Smith GH. Reprogramming human cancer cells in the mouse mammary gland. Cancer Res. 2010;70:6336–43.PubMedCrossRefGoogle Scholar
  65. 65.
    Booth BW, Boulanger CA, Anderson LH, Smith GH. The normal mammary microenvironment suppresses the tumorigenic phenotype of mouse mammary tumor virus-neu-transformed mammary tumor cells. Oncogene. 2011;30:679–89.PubMedCrossRefGoogle Scholar
  66. 66.
    Kasemeier-Kulesa JC, Teddy JM, Postovit LM, et al. Reprogramming multipotent tumor cells with the embryonic neural crest microenvironment. Dev Dyn. 2008;237:2657–66.PubMedCrossRefGoogle Scholar
  67. 67.
    Postovit LM, Margaryan NV, Seftor EA, et al. Human embryonic stem cell microenvironment suppresses the tumorigenic phenotype of aggressive cancer cells. Proc Natl Acad Sci U S A. 2008;105: 4329–34.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science + Business Media New York 2013

Authors and Affiliations

  • Brian W. Booth
    • 1
  • Daniel Medina
    • 2
  • Gilbert H. Smith
    • 3
  1. 1.Rhodes Engineering Research Center, Institute for Biological Interfaces of EngineeringClemson UniversityClemsonUSA
  2. 2.Department of Molecular and Cellular BiologyBaylor College of MedicineHoustonUSA
  3. 3.Cell and Cancer BiologyNational Cancer InstituteBethesdaUSA

Personalised recommendations