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Three-Dimensional Cultures of Mouse Mammary Epithelial Cells

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Epithelial Cell Culture Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 945))

Abstract

The mammary gland is an ideal “model organism” for studying tissue specificity and gene expression in mammals: it is one of the few organs that develop after birth and it undergoes multiple cycles of growth, differentiation and regression during the animal’s lifetime in preparation for the important function of lactation. The basic “functional differentiation” unit in the gland is the mammary acinus made up of a layer of polarized epithelial cells specialized for milk production surrounded by myoepithelial contractile cells, and the two-layered structure is surrounded by basement membrane. Much knowledge about the regulation of mammary gland development has been acquired from studying the physiology of the gland and of lactation in rodents. Culture studies, however, were hampered by the inability to maintain functional differentiation on conventional tissue culture plastic. We now know that the microenvironment, including the extracellular matrix and tissue architecture, plays a crucial role in directing functional differentiation of organs. Thus, in order for culture systems to be effective experimental models, they need to recapitulate the basic unit of differentiated function in the tissue or organ and to maintain its three-dimensional (3D) structure. Mouse mammary culture models evolved from basic monolayers of cells to an array of complex 3D systems that observe the importance of the microenvironment in dictating proper tissue function and structure. In this chapter, we focus on how 3D mouse mammary epithelial cultures have enabled investigators to gain a better understanding of the organization, development and function of the acinus, and to identify key molecular, structural, and mechanical cues important for maintaining mammary function and architecture. The accompanying chapter of Vidi et al. describes 3D models developed for human cells. Here, we describe how mouse primary epithelial cells and cell lines—essentially those we use in our laboratory—are cultured in relevant 3D microenvironments. We focus on the design of functional assays that enable us to understand the intricate signaling events underlying mammary gland biology, and address the advantages and limitations of the different culture settings. Finally we also discuss how advances in bioengineering tools may help towards the ultimate goal of building tissues and organs in culture for basic research and clinical studies.

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Acknowledgments

We thank Dr. Joni Mott for critical reading of the manuscript and Dr. Hidetoshi Mori for providing images of cells in Fig. 2b, c. The work from MJB’s laboratory is supported by grants from the US Department of Energy, Office of Biological and Environmental Research, a Distinguished Fellow Award to MJB and Low Dose Radiation Program (contract no. DE-AC02-05CH1123); by National Cancer Institute (awards R37CA064786, U54CA126552, R01CA057621, U54CA112970, U54CA143836, U01CA143233); and by US Department of Defense (W81XWH0810736). RM is supported by US Department of Defense, BCRP fellowship (W81XWH0810481).

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  1. Lawrence Berkeley National Laboratory, Berkeley, CA, USA

    Rana Mroue & Mina J. Bissell

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  1. Rana Mroue
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  2. Mina J. Bissell
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Correspondence to Mina J. Bissell .

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  1. Cystic Fibrosis/Pulmonary Research Ctr., University of North Carolina, Thurston Bowles Building 4011, Chapel Hill, 27599, North Carolina, USA

    Scott H. Randell

  2. Cystic Fibrosis/Pulmonary Research Ctr., The University of North Carolina, Thurston-Bowles Building 4011, Chapel Hill, 27599, North Carolina, USA

    M. Leslie Fulcher

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Mroue, R., Bissell, M.J. (2012). Three-Dimensional Cultures of Mouse Mammary Epithelial Cells. In: Randell, S., Fulcher, M. (eds) Epithelial Cell Culture Protocols. Methods in Molecular Biology, vol 945. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-125-7_14

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