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Reconstruction of human mammary tissues in a mouse model


Establishing a model system that more accurately recapitulates both normal and neoplastic breast epithelial development in rodents is central to studying human breast carcinogenesis. However, the inability of human breast epithelial cells to colonize mouse mammary fat pads is problematic. Considering that the human breast is a more fibrous tissue than is the adipose-rich stroma of the murine mammary gland, our group sought to bypass the effects of the rodent microenvironment through incorporation of human stromal fibroblasts. We have been successful in reproducibly recreating functionally normal breast tissues from reduction mammoplasty tissues, in what we term the human-in-mouse (HIM) model. Here we describe our relatively simple and inexpensive techniques for generating this orthotopic xenograft model. Whether the model is to be applied for understanding normal human breast development or tumorigenesis, investigators with minimal animal surgery skills, basic cell culture techniques and access to human breast tissue will be able to generate humanized mouse glands within 3 months. Clearing the mouse of its endogenous epithelium with subsequent stromal humanization takes 1 month. The subsequent implantation of co-mixed human epithelial cells and stromal cells occurs 2 weeks after humanization, so investigators should expect to observe the desired outgrowths 2 months afterward. As a whole, this model system has the potential to improve the understanding of crosstalk between tissue stroma and the epithelium as well as factors involved in breast stem cell biology tumor initiation and progression.

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Figure 1: Flowchart and timeline of the HIM protocol.
Figure 3: Formation of ductal, lobular and acinar structures in HIM model.
Figure 2: Lentiviral-mediated transduction of GFP into human breast organoids.


  1. DeOme, K.B., Faulkin, L.J., Jr., Bern, H. & Blair, P.B. Development of mammary tumors from hyperplastic alveolar nodules transplanted into gland-free mammary fat pads of female C3H Mice. Cancer Res 19, 515–525 (1959).

    CAS  PubMed  Google Scholar 

  2. Daniel, C.W. & DeOme, K.B. Growth of mouse mammary glands in vivo after monolayer culture. Science 149, 634–636 (1965).

    Article  CAS  PubMed  Google Scholar 

  3. Young, L.J., Medina, D., DeOme, K.B. & Daniel, C.W. The influence of host and tissue age on life span and growth rate of serially transplanted mouse mammary gland. Exp. Gerontol. 6, 49–56 (1971).

    Article  CAS  PubMed  Google Scholar 

  4. Medina, D. & DeOme, K.B. Carcinogen-induced mammary tumors from preneoplastic nodule outgrowths in BALB-c mice. Cancer Res. 30, 1055–1059 (1970).

    CAS  PubMed  Google Scholar 

  5. Outzen, H.C. & Custer, R.P. Growth of human normal and neoplastic mammary tissues in the cleared fat pad of the nude mouse. J. Natl. Cancer Inst. 55, 1461–1466 (1975).

    Article  CAS  PubMed  Google Scholar 

  6. Sheffield, L.G. & Welsch, C.W. Transplantation of human breast epithelia to mammary-gland-free fat-pads of athymic nude mice: influnence of mammatrophic hormones of breast epithelia. Int. J. Cancer 41, 713–719 (1988).

    Article  CAS  PubMed  Google Scholar 

  7. Parmar, H. et al. A novel method for growing human breast epithelium in vivo using mouse and human mammary fibroblasts. Endocrinology 143, 4886–4896 (2002).

    Article  CAS  PubMed  Google Scholar 

  8. Kuperwasser, C. et al. Reconstruction of functionally normal and malignant human breast tissues in mice. Proc. Natl. Acad. Sci. U.S.A. 101, 4966–4971 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Rebar, R.W., Morandini, I.C., Petze, J.E. & Erickson, G.F. The hormonal basis of reproductive defects in athymic mice: diminished gonadotropin concentration in prepubertal females. Endocrinology 108, 120–126 (1981).

    Article  CAS  PubMed  Google Scholar 

  10. Young, L.J.T. Methods in Mammary Gland Biology and Breast Cancer Research. Ip MMaABB (ed.), pp. 67–74 (Kluwer Academic/Plenum, New York, 2000).

    Book  Google Scholar 

  11. Barcellos-Hoff, M.H. & Ravani, S.A. Irradiated mammary gland stroma promotes the expression of tumorigenic potential by unirradiated epithelial cells. Cancer Res. 60, 1254–1260 (2000).

    CAS  PubMed  Google Scholar 

  12. Ohuchida, K. et al. Radiation to stromal fibroblasts increases invasiveness of pancreatic cancer cells through tumor-stromal interactions. Cancer Res. 64, 3215–3222 (2004).

    Article  CAS  PubMed  Google Scholar 

  13. Tsai, K.K.C., Chuang, E.Y.-Y., Little, J.B. & Yuan, Z.M. Cellular mechanisms for low-dose ionizing radiation-induced perturbation of the breast tissue microenvironment. Cancer Res. 65, 6734–6744 (2005).

    Article  CAS  PubMed  Google Scholar 

  14. Rasmussen, S.B., Young, L.J.T. & Smith, G.H. Methods in Mammary Gland Biology and Breast Cancer Research. Ip MMaABB (ed.), pp. 75–86 (Kluwer Academic/Plenum, New York, 2000).

    Book  Google Scholar 

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This work was supported by grants from the RB Sackler and Susan Komen Foundations. C.K. is an RB Sackler Scholar.

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Correspondence to Charlotte Kuperwasser.

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Kuperwasser is a consultant of AVEO Pharmaceuticals Inc. Charlotte Kuperwasser has filed a patent application based on the work reported in this manuscript.

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Proia, D., Kuperwasser, C. Reconstruction of human mammary tissues in a mouse model. Nat Protoc 1, 206–214 (2006).

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