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Reconstruction of human mammary tissues in a mouse model

Abstract

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.

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Acknowledgements

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). https://doi.org/10.1038/nprot.2006.31

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