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3D Culture Assays of Murine Mammary Branching Morphogenesis and Epithelial Invasion

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Tissue Morphogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1189))

Abstract

Epithelia are fundamental tissues that line cavities, glands, and outer body surfaces. We use three-dimensional (3D) embedded culture of primary murine mammary epithelial ducts, called “organoids,” to recapitulate in days in culture epithelial programs that occur over weeks deep within the body. Modulating the composition of the extracellular matrix (ECM) allows us to model cell- and tissue-level behaviors observed in normal development, such as branching morphogenesis, and in cancer, such as invasion and dissemination. Here, we describe a collection of protocols for 3D culture of mammary organoids in different ECMs and for immunofluorescence staining of 3D culture samples and mammary gland tissue sections. We illustrate expected phenotypic outcomes of each assay and provide troubleshooting tips for commonly encountered technical problems.

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Acknowledgements

A.J.E. was supported by a Research Scholar Grant, RSG-12-141-01-CSM from the American Cancer Society, E.R.S. was supported by the Isaac Morris Hay and Lucille Elizabeth Hay Graduate Fellowship Award, R.J.H. was supported by an NIH/NIGMS training grant (2T32GM007445), and K.J.C. was supported by the US Department of Defense (W81XWH-12-1-0018).

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Authors and Affiliations

  1. Departments of Cell Biology and Oncology, Center for Cell Dynamics, Johns Hopkins School of Medicine, 855 N. Wolfe Street, 452 Rangos Building, Baltimore, MD, 21205, USA

    Kim-Vy Nguyen-Ngoc, Eliah R. Shamir, Robert J. Huebner, Jennifer N. Beck, Kevin J. Cheung & Andrew J. Ewald Ph.D.

Authors
  1. Kim-Vy Nguyen-Ngoc
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  2. Eliah R. Shamir
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  3. Robert J. Huebner
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  4. Jennifer N. Beck
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  5. Kevin J. Cheung
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  6. Andrew J. Ewald Ph.D.
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Corresponding author

Correspondence to Andrew J. Ewald Ph.D. .

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Editors and Affiliations

  1. Chemical & Biological Engineering, Princeton University, New Jersey, USA

    Celeste M. Nelson

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Nguyen-Ngoc, KV., Shamir, E.R., Huebner, R.J., Beck, J.N., Cheung, K.J., Ewald, A.J. (2015). 3D Culture Assays of Murine Mammary Branching Morphogenesis and Epithelial Invasion. In: Nelson, C. (eds) Tissue Morphogenesis. Methods in Molecular Biology, vol 1189. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1164-6_10

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  • DOI: https://doi.org/10.1007/978-1-4939-1164-6_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1163-9

  • Online ISBN: 978-1-4939-1164-6

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