Skip to main content
SpringerLink
Log in

Analysis of Mammary Gland Phenotypes by Transplantation of the Genetically Marked Mammary Epithelium

  • Protocol
  • First Online:
Book cover Mammary Gland Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1501))

Abstract

The mammary gland is the only organ to undergo most of its development after birth and therefore particularly attractive for studying developmental processes. In the mouse, powerful tissue recombination techniques are available that can be elegantly combined with the use of different genetically engineered mouse models to study development and differentiation in vivo.

In this chapter, we describe how epithelial intrinsic gene function can by discerned by grafting mammary epithelial cells of different genotypes to wild-type recipients. Either pieces of mammary epithelial tissue or dissociated mammary epithelial cells are isolated from donor mice and subsequently transplanted into recipients whose mammary fat pads were divested of their endogenous epithelium. This is followed by phenotypic characterization of the epithelial outgrowth either by fluorescence stereomicroscopy for the fluorescently marked grafts or carmine alum whole mount for the unmarked epithelia.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Knight CH, Peaker M (1982) Development of the mammary gland. J Reprod Fertil 65:521–536

    Article  CAS  PubMed  Google Scholar 

  2. Faulkin LJ, DeOme KB (1958) The Effect of estradiol and cortisol on the transplantability and subsequent fate of normal, hyperplastic, and tumorous mammary tissue of C3H Mice. Cancer Res 18:51–56

    CAS  Google Scholar 

  3. Daniel CW, Deome KB (1965) Growth of mouse mammary glands in vivo after monolayer culture. Science 149:634–636

    Article  CAS  PubMed  Google Scholar 

  4. Deome KB, Faulkin LJ Jr, Bern HA, Blair PB (1959) Development of mammary tumors from hyperplastic alveolar nodules transplanted into gland-free mammary fat pads of female C3H mice. Cancer Res 19:515–520

    CAS  PubMed  Google Scholar 

  5. Hoshino K, Gardner WU, Pawlikowski RA (1965) The incidence of cancer in quantitatively transplanted mammary glands and its relation to age and milk agent of the donor and host mice. Cancer Res 25:1792–1803

    CAS  PubMed  Google Scholar 

  6. Muhlbock O (1956) The hormonal genesis of mammary cancer. Adv Cancer Res 4:371–391

    Article  CAS  PubMed  Google Scholar 

  7. Prehn RT (1953) Tumors and hyperplastic nodules in transplanted mammary glands. J Natl Cancer Inst 13:859–871

    CAS  PubMed  Google Scholar 

  8. Shimkin MB, Wyman RS, Andervont HB (1946) Mammary tumors in mice following transplantation of mammary tissue. J Natl Cancer Inst 7:77

    CAS  PubMed  Google Scholar 

  9. Hoshino K (1962) Morphogenesis and growth potentiality of mammary glands in mice. I. Transplantability and growth potentiality of mammary tissue of virgin mice. J Natl Cancer Inst 29:835–851

    CAS  PubMed  Google Scholar 

  10. Hoshino K (1963) Morphogenesis and growth potentiality of mammary glands in mice. II. Quantitative transplantation of mammary glands of normal male mice. J Natl Cancer Inst 30:585–591

    CAS  PubMed  Google Scholar 

  11. Hoshino K (1964) Regeneration and growth of quantitatively transplanted mammary glands of normal female mice. Anat Rec 150:221–235

    Article  CAS  PubMed  Google Scholar 

  12. Hoshino K (1967) Transplantability of mammary gland in brown fat pads of mice. Nature 213:194–195

    Article  CAS  PubMed  Google Scholar 

  13. Hoshino K, Gardner WU (1967) Transplantability and life span of mammary gland during serial transplantation in mice. Nature 213:193–194

    Article  CAS  PubMed  Google Scholar 

  14. Brisken C, Park S, Vass T, Lydon JP, O'Malley BW, Weinberg RA (1998) A paracrine role for the epithelial progesterone receptor in mammary gland development. Proc Natl Acad Sci U S A 95:5076–5081

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Mallepell S, Krust A, Chambon P, Brisken C (2006) 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 103:2196–2201

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Ciarloni L, Mallepell S, Brisken C (2007) Amphiregulin is an essential mediator of estrogen receptor alpha function in mammary gland development. Proc Natl Acad Sci U S A 104:5455–5460

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Brisken C, Heineman A, Chavarria T, Elenbaas B, Tan J, Dey SK et al (2000) Essential function of Wnt-4 in mammary gland development downstream of progesterone signaling. Genes Dev 14:650–654

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Heckman-Stoddard BM, Vargo-Gogola T, Herrick MP, Visbal AP, Lewis MT, Settleman J et al (2011) P190A RhoGAP is required for mammary gland development. Dev Biol 360:1–10

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Pond AC, Bin X, Batts T, Roarty K, Hilsenbeck S, Rosen JM (2013) Fibroblast growth factor receptor signaling is essential for normal mammary gland development and stem cell function. Stem Cells 31:178–189

    Article  CAS  PubMed  Google Scholar 

  20. Shackleton M, Vaillant F, Simpson KJ, Stingl J, Smyth GK, Asselin-Labat ML et al (2006) Generation of a functional mammary gland from a single stem cell. Nature 439:84–88

    Article  CAS  PubMed  Google Scholar 

  21. Sleeman KE, Kendrick H, Robertson D, Isacke CM, Ashworth A, Smalley MJ (2007) Dissociation of estrogen receptor expression and in vivo stem cell activity in the mammary gland. J Cell Biol 176:19–26

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Friedrich G, Soriano P (1991) Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice. Genes Dev 5:1513–1523

    Article  CAS  PubMed  Google Scholar 

  23. Okabe M, Ikawa M, Kominami K, Nakanishi T, Nishimune Y (1997) 'Green mice' as a source of ubiquitous green cells. FEBS Lett 407:313–319

    Article  CAS  PubMed  Google Scholar 

  24. Vintersten K, Monetti C, Gertsenstein M, Zhang P, Laszlo L, Biechele S et al (2004) Mouse in red: red fluorescent protein expression in mouse ES cells, embryos, and adult animals. Genesis 40:241–246

    Article  CAS  PubMed  Google Scholar 

  25. Smalley MJ (2010) Isolation, culture and analysis of mouse mammary epithelial cells. Methods Mol Biol 633:139–170

    Article  CAS  PubMed  Google Scholar 

  26. LaMarca HL, Visbal AP, Creighton CJ, Liu H, Zhang Y, Behbod F et al (2010) CCAAT/enhancer binding protein beta regulates stem cell activity and specifies luminal cell fate in the mammary gland. Stem Cells 28:535–544

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Daniel CW, Deome KB, Young JT, Blair PB, Faulkin LJ Jr (2009) The in vivo life span of normal and preneoplastic mouse mammary glands: a serial transplantation study. J Mammary Gland Biol Neoplasia 14:355–362

    Article  PubMed  Google Scholar 

  28. Faulkin LJ Jr, Deome KB (1960) Regulation of growth and spacing of gland elements in the mammary fat pad of the C3H mouse. J Natl Cancer Inst 24:953–969

    PubMed  Google Scholar 

  29. Williams MF, Hoshino K (1970) Early histogenesis of transplanted mouse mammary glands. I. Within 21 days following isografting. Z Anat Entwicklungsgesch 132:305–317

    Article  CAS  PubMed  Google Scholar 

  30. Flanagan SP (1966) 'Nude', a new hairless gene with pleiotropic effects in the mouse. Genet Res 8:295–309

    Article  CAS  PubMed  Google Scholar 

  31. Seibert K, Shafie SM, Triche TJ, Whang-Peng JJ, O'Brien SJ, Toney JH et al (1983) Clonal variation of MCF-7 breast cancer cells in vitro and in athymic nude mice. Cancer Res 43:2223–2239

    CAS  PubMed  Google Scholar 

  32. Soule HD, McGrath CM (1980) Estrogen responsive proliferation of clonal human breast carcinoma cells in athymic mice. Cancer Lett 10:177–189

    Article  CAS  PubMed  Google Scholar 

  33. Mombaerts P, Iacomini J, Johnson RS, Herrup K, Tonegawa S, Papaioannou VE (1992) RAG-1-deficient mice have no mature B and T lymphocytes. Cell 68:869–877

    Article  CAS  PubMed  Google Scholar 

  34. Shinkai Y, Rathbun G, Lam KP, Oltz EM, Stewart V, Mendelsohn M et al (1992) RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D)J rearrangement. Cell 68:855–867

    Article  CAS  PubMed  Google Scholar 

  35. Reed JR, Schwertfeger KL (2010) Immune cell location and function during post-natal mammary gland development. J Mammary Gland Biol Neoplasia 15:329–339

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors thank all current and former members of the Brisken laboratory, who contributed to the development of these techniques, and Gisèle Ferrand for advice on anesthesia procedures.

Author information

Authors and Affiliations

  1. The Swiss Institute for Experimental Cancer Research (ISREC), Swiss Federal Institute of Technology (EPFL), Station 19, CH-1015, Lausanne, Switzerland

    Duje Buric & Cathrin Brisken

Authors
  1. Duje Buric
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Cathrin Brisken
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Cathrin Brisken .

Editor information

Editors and Affiliations

  1. School of Biomolecular and Biomedical Science, University College Dublin, Belfield, Dublin, Ireland

    Finian Martin

  2. Institute of Cancer Sciences College of MVLS University of Glasgow, Glasgow, United Kingdom

    Torsten Stein

  3. Division of Oncology-Pathology, Lund University Cancer Center/Medicon Village, Lund, Sweden

    Jillian Howlin

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media New York

About this protocol

Cite this protocol

Buric, D., Brisken, C. (2017). Analysis of Mammary Gland Phenotypes by Transplantation of the Genetically Marked Mammary Epithelium. In: Martin, F., Stein, T., Howlin, J. (eds) Mammary Gland Development. Methods in Molecular Biology, vol 1501. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6475-8_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6475-8_4

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6473-4

  • Online ISBN: 978-1-4939-6475-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation