Skip to main content

Advertisement

SpringerLink
Log in

Enrichment for Repopulating Cells and Identification of Differentiation Markers in the Bovine Mammary Gland

  • Published:
Journal of Mammary Gland Biology and Neoplasia Aims and scope Submit manuscript

Abstract

Elucidating cell hierarchy in the mammary gland is fundamental for understanding the mechanisms governing its normal development and malignant transformation. There is relatively little information on cell hierarchy in the bovine mammary gland, despite its agricultural potential and relevance to breast cancer research. Challenges in bovine-to-mouse xenotransplantation and difficulties obtaining bovine-compatible antibodies hinder the study of mammary stem-cell dynamics in this species. In-vitro indications of distinct bovine mammary epithelial cell populations, sorted according to CD24 and CD49f expression, have been provided. Here, we successfully transplanted these bovine populations into the cleared fat pads of immunocompromised mice, providing in-vivo evidence for the multipotency and self-renewal capabilities of cells that are at the top of the cell hierarchy (termed mammary repopulating units). Additional outgrowths from transplantation, composed exclusively of myoepithelial cells, were indicative of unipotent basal stem cells or committed progenitors. Sorting luminal cells according to E-cadherin revealed three distinct populations: luminal progenitors, and early- and late-differentiating cells. Finally, miR-200c expression was negatively correlated with differentiation levels in both the luminal and basal branches of the bovine mammary cell hierarchy. Together, these experiments provide further evidence for the presence of a regenerative entity in the bovine mammary gland and for the multistage differentiation process within the luminal lineage.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Shackleton M, Vaillant F, Simpson KJ, Stingl J, Smyth GK, Asselin-Labat ML, et al. Generation of a functional mammary gland from a single stem cell. Nature. 2006;439(7072):84–8.

    Article  CAS  PubMed  Google Scholar 

  2. Stingl J, Eirew P, Ricketson I, Shackleton M, Vaillant F, Choi D, et al. Purification and unique properties of mammary epithelial stem cells. Nature. 2006;439(7079):993–7.

    CAS  PubMed  Google Scholar 

  3. Rios AC, Fu NY, Lindeman GJ, Visvader JE. In situ identification of bipotent stem cells in the mammary gland. Nature. 2014;506(7488):322–7.

    Article  CAS  PubMed  Google Scholar 

  4. Capuco AV, Ellis SE. Comparative aspects of mammary gland development and homeostasis. Annu Rev Anim Biosci. 2013;1:179–202.

    Article  PubMed  Google Scholar 

  5. 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(1):53–68.

    Article  CAS  PubMed  Google Scholar 

  6. Rauner G, Barash I. Cell hierarchy and lineage commitment in the bovine mammary gland. PLoS One. 2012;7(1):e30113.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. 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(5):515–20.

    CAS  PubMed  Google Scholar 

  8. 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(1):21–31.

    Article  CAS  PubMed  Google Scholar 

  9. Sheffield LG. Organization and growth of mammary epithelia in the mammary gland fat pad. J Dairy Sci. 1988;71(10):2855–74.

    Article  CAS  PubMed  Google Scholar 

  10. Rauner G, Leviav A, Mavor E, Barash I. Development of foreign mammary epithelial morphology in the stroma of immunodeficient mice. PLoS One. 2013;8(6):e68637.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Pece S, Gutkind JS. Signaling from E-cadherins to the MAPK pathway by the recruitment and activation of epidermal growth factor receptors upon cell-cell contact formation. J Biol Chem. 2000;275(52):41227–33.

    Article  CAS  PubMed  Google Scholar 

  12. Fedor-Chaiken M, Hein PW, Stewart JC, Brackenbury R, Kinch MS. E-cadherin binding modulates EGF receptor activation. Cell Commun Adhes. 2003;10(2):105–18.

    Article  CAS  PubMed  Google Scholar 

  13. Boussadia O, Kutsch S, Hierholzer A, Delmas V, Kemler R. E-cadherin is a survival factor for the lactating mouse mammary gland. Mech Dev. 2002;115(1–2):53–62.

    Article  CAS  PubMed  Google Scholar 

  14. Shamir ER, Pappalardo E, Jorgens DM, Coutinho K, Tsai WT, Aziz K, et al. Twist1-induced dissemination preserves epithelial identity and requires E-cadherin. J Cell Biol. 2014;204(5):839–56.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Hilmarsdottir B, Briem E, Bergthorsson JT, Magnusson MK, Gudjonsson T. Functional role of the microRNA-200 family in breast morphogenesis and neoplasia. Genes (Basel). 2014;5(3):804–20.

    Google Scholar 

  16. Shimono Y, Zabala M, Cho RW, Lobo N, Dalerba P, Qian D, et al. Downregulation of miRNA-200c links breast cancer stem cells with normal stem cells. Cell. 2009;138(3):592–603.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Schmalhofer O, Brabletz S, Brabletz T. E-cadherin, beta-catenin, and ZEB1 in malignant progression of cancer. Cancer Metastasis Rev. 2009;28(1–2):151–66.

    Article  CAS  PubMed  Google Scholar 

  18. 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(5):713–9.

    Article  CAS  PubMed  Google Scholar 

  19. Kuperwasser C, Chavarria T, Wu M, Magrane G, Gray JW, Carey L, et al. Reconstruction of functionally normal and malignant human breast tissues in mice. Proc Natl Acad Sci U S A. 2004;101(14):4966–71.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Sheffield LG, Welsch CW. Transplantation of bovine mammary tissue to athymic nude mice: growth subcutaneously and in mammary gland-free fat pads. J Dairy Sci. 1986;69(4):1141–7.

    Article  CAS  PubMed  Google Scholar 

  21. Martignani E, Eirew P, Accornero P, Eaves CJ, Baratta M. Human milk protein production in xenografts of genetically engineered bovine mammary epithelial stem cells. PLoS One. 2010;5(10):e13372.

    Article  PubMed  PubMed Central  Google Scholar 

  22. Williams JM, Daniel CW. Mammary ductal elongation: differentiation of myoepithelium and basal lamina during branching morphogenesis. Dev Biol. 1983;97(2):274–90.

    Article  CAS  PubMed  Google Scholar 

  23. Gudjonsson T, Ronnov-Jessen L, Villadsen R, Rank F, Bissell MJ, Petersen OW. Normal and tumor-derived myoepithelial cells differ in their ability to interact with luminal breast epithelial cells for polarity and basement membrane deposition. J Cell Sci. 2002;115(Pt 1):39–50.

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Shehata M, Teschendorff A, Sharp G, Novcic N, Russell IA, Avril S, et al. Phenotypic and functional characterisation of the luminal cell hierarchy of the mammary gland. Breast Cancer Res. 2012;14(5):R134.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Locke D, Perusinghe N, Newman T, Jayatilake H, Evans WH, Monaghan P. Developmental expression and assembly of connexins into homomeric and heteromeric gap junction hemichannels in the mouse mammary gland. J Cell Physiol. 2000;183(2):228–37.

    Article  CAS  PubMed  Google Scholar 

  26. Asselin-Labat ML, Sutherland KD, Barker H, Thomas R, Shackleton M, Forrest NC, et al. Gata-3 is an essential regulator of mammary-gland morphogenesis and luminal-cell differentiation. Nat Cell Biol. 2007;9(2):201–9.

    Article  CAS  PubMed  Google Scholar 

  27. Gregory PA, Bert AG, Paterson EL, Barry SC, Tsykin A, Farshid G, et al. The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol. 2008;10(5):593–601.

    Article  CAS  PubMed  Google Scholar 

  28. Park IK, Qian D, Kiel M, Becker MW, Pihalja M, Weissman IL, et al. Bmi-1 is required for maintenance of adult self-renewing haematopoietic stem cells. Nature. 2003;423(6937):302–5.

    Article  CAS  PubMed  Google Scholar 

  29. Bockmeyer CL, Christgen M, Muller M, Fischer S, Ahrens P, Langer F, et al. MicroRNA profiles of healthy basal and luminal mammary epithelial cells are distinct and reflected in different breast cancer subtypes. Breast Cancer Res Treat. 2011;130(3):735–45.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Animal Science, ARO, The Volcani Center, Bet-Dagan, Israel

    Gat Rauner & Itamar Barash

  2. The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Jerusalem, Israel

    Gat Rauner

Authors
  1. Gat Rauner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Itamar Barash
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Itamar Barash.

Ethics declarations

Funding

Grants from the Israel Science Foundation, Israel Academy of Sciences, contract number: 289/11 to IB, ChiefScientist, Israeli Ministry of Agriculture and the Israeli Milk Marketing Board to IB.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Rauner, G., Barash, I. Enrichment for Repopulating Cells and Identification of Differentiation Markers in the Bovine Mammary Gland. J Mammary Gland Biol Neoplasia 21, 41–49 (2016). https://doi.org/10.1007/s10911-015-9348-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10911-015-9348-x

Keywords

Search

Navigation