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Three-dimensional Mammary Epithelial Cell Morphogenesis Model for Analysis of TGFß Signaling

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TGF-β Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1344))

Abstract

Culturing mammary epithelial cells in laminin-rich extracellular matrices (three dimensional or 3D culture) offers significant advantages over that in the conventional two-dimensional (2D) tissue culture system in that it takes into considetation the impact of extracellular matrix (ECM) microenvironment on the proliferation, survival, and differentiation of mammary epithelial cells. When grown in the 3D culture, untransformed mammary epithelial cells undergo morphogenesis to form a multicellular and polarized acini-like structure that functionally mimics the differentiated alveoli in the pregnancy mammary gland. This process is subjected to regulation by many growth factors and cytokines. The transforming growth factor-ß (TGFß) is a multipotent cytokine that regulates multiple aspects of development and tumorigenesis. In addition to its effects on epithelial cell proliferation, survival, and differentiation, it is also a potent regulator of the cell–matrix interaction. Thus, the 3D culture model may recapitulate the complex in vivo epithelial cell microenvironment and allow us to fully evaluate the role of TGFß signaling in multiple aspects of normal and cancerous cell behavior. In this chapter we provide detailed protocols for growing mammary epithelial cells in the 3D Matrigel for analysis of signaling pathways.

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Acknowledgement

We would like to thank members of Dr. Mina Bissell’s group for their assistance. The work is supported by NIH RO1 CA101891 and RO1 DK090347 to K. Luo. J. Rashidian is supported by the Susan G. Komen For The Cure KG101263.

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Authors and Affiliations

  1. Department of Molecular and Cell Biology (MCB), University of California, 16 Barker Hall # 3204, Berkeley, CA, 94720-3204, USA

    Juliet Rashidian & Kunxin Luo Ph.D.

Authors
  1. Juliet Rashidian
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  2. Kunxin Luo Ph.D.
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Corresponding author

Correspondence to Kunxin Luo Ph.D. .

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Editors and Affiliations

  1. Departments of Surgery and Molecular & Cellular Biology, Baylor College of Medicine, Houston, Texas, USA

    Xin-Hua Feng

  2. Life Sciences Institute, Zhejiang University, Hangzhou, Zhejiang Proviince, China

    Pinglong Xu

  3. Department of Surgery, Baylor College of Medicine, Houston, Texas, USA

    Xia Lin

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Rashidian, J., Luo, K. (2016). Three-dimensional Mammary Epithelial Cell Morphogenesis Model for Analysis of TGFß Signaling. In: Feng, XH., Xu, P., Lin, X. (eds) TGF-β Signaling. Methods in Molecular Biology, vol 1344. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2966-5_7

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  • DOI: https://doi.org/10.1007/978-1-4939-2966-5_7

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2965-8

  • Online ISBN: 978-1-4939-2966-5

  • eBook Packages: Springer Protocols

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