Abstract
Epithelial–stromal interactions play an essential role in regulation of mammary gland development, homeostasis, and tumorigenesis. Fibroblasts constitute a substantial proportion of mammary gland stromal cells in human breast and have been recognized for their paracrine signaling and extracellular matrix production and remodeling roles during normal breast development as well as in breast cancer. However, our current knowledge on human breast fibroblast functions is incomplete. Here we provide a detailed protocol for an organotypic human breast assay to facilitate research in the roles of human breast fibroblasts in mammary epithelial morphogenesis and early tumorigenesis.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
McNally S, Stein T (2017) Overview of mammary gland development: a comparison of mouse and human, methods Mol. Biol 1501:1–17. https://doi.org/10.1007/978-1-4939-6475-8_1
Gjorevski N, Nelson CM (2011) Integrated morphodynamic signalling of the mammary gland. Nat Rev Mol Cell Biol 12:581–593. https://doi.org/10.1038/nrm3168
Sumbal J, Belisova D, Koledova Z (2020) Fibroblasts: the grey eminence of mammary gland development. Semin Cell Dev Biol 111:134–142. https://doi.org/10.1016/j.semcdb.2020.10.012
Koledova Z, Zhang X, Streuli C, Clarke RB, Klein OD, Werb Z, Lu P (2016) SPRY1 regulates mammary epithelial morphogenesis by modulating EGFR-dependent stromal paracrine signaling and ECM remodeling. Proc Natl Acad Sci U S A 113:E5731–E5740. https://doi.org/10.1073/pnas.1611532113
Sumbal J, Koledova Z (2019) FGF signaling in mammary gland fibroblasts regulates multiple fibroblast functions and mammary epithelial morphogenesis. Development 146(23):dev185306. https://doi.org/10.1242/dev.185306
Peuhu E, Kaukonen R, Lerche M, Saari M, Guzmán C, Rantakari P, De Franceschi N, Wärri A, Georgiadou M, Jacquemet G, Mattila E, Virtakoivu R, Liu Y, Attieh Y, Silva KA, Betz T, Sundberg JP, Salmi M, Deugnier M-A, Eliceiri KW, Ivaska J (2017) SHARPIN regulates collagen architecture and ductal outgrowth in the developing mouse mammary gland. EMBO J 36:165–182. https://doi.org/10.15252/embj.201694387
Hammer AM, Sizemore GM, Shukla VC, Avendano A, Sizemore ST, Chang JJ, Kladney RD, Cuitiño MC, Thies KA, Verfurth Q, Chakravarti A, Yee LD, Leone G, Song JW, Ghadiali SN, Ostrowski MC (2017) Stromal PDGFR-α activation enhances matrix stiffness, impedes mammary ductal development, and accelerates tumor growth. Neoplasia 19:496–508. https://doi.org/10.1016/j.neo.2017.04.004
Houthuijzen JM, Jonkers J (2018) Cancer-associated fibroblasts as key regulators of the breast cancer tumor microenvironment. Cancer Metastasis Rev 37:577–597. https://doi.org/10.1007/s10555-018-9768-3
Gudjonsson T, Villadsen R, Nielsen HL, Rønnov-Jessen L, Bissell MJ, Petersen OW (2002) Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties. Genes Dev 16:693–706. https://doi.org/10.1101/gad.952602
Briem E, Ingthorsson S, Traustadottir GA, Hilmarsdottir B, Gudjonsson T (2019) Application of the D492 cell lines to explore breast morphogenesis, EMT and cancer progression in 3D culture. J Mammary Gland Biol Neoplasia 24:139–147. https://doi.org/10.1007/s10911-018-09424-w
Morera E, Steinhäuser SS, Budkova Z, Ingthorsson S, Kricker J, Krueger A, Traustadottir GA, Gudjonsson T (2019) YKL-40/CHI3L1 facilitates migration and invasion in HER2 overexpressing breast epithelial progenitor cells and generates a niche for capillary-like network formation. In Vitro Cell Dev Biol Anim 55:838–853. https://doi.org/10.1007/s11626-019-00403-x
Steinhaeuser SS, Morera E, Budkova Z, Schepsky A, Wang Q, Rolfsson O, Riedel A, Krueger A, Hilmarsdottir B, Maelandsmo GM, Valdimarsdottir B, Sigurdardottir AK, Agnarsson BA, Jonasson JG, Ingthorsson S, Traustadottir GA, Oskarsson T, Gudjonsson T (2020) ECM1 secreted by HER2-overexpressing breast cancer cells promotes formation of a vascular niche accelerating cancer cell migration and invasion. Lab Investig 100:928–944. https://doi.org/10.1038/s41374-020-0415-6
Koledova Z (2017) 3D coculture of mammary organoids with Fibrospheres: a model for studying epithelial-stromal interactions during mammary branching morphogenesis. Methods Mol Biol 1612:107–124. https://doi.org/10.1007/978-1-4939-7021-6_8
Koledova Z, Lu P (2017) A 3D fibroblast-epithelium co-culture model for understanding microenvironmental role in branching morphogenesis of the mammary gland. Methods Mol Biol 1501:217–231. https://doi.org/10.1007/978-1-4939-6475-8_10
Acknowledgments
We thank present and past members of our laboratories for their technical support. This work was supported by the Icelandic Cancer Society Science fund (G.A.T.), the Grant Agency of Masaryk University (projects no. MUNI/G/1446/2018 to Z.K.), and funds from the Faculty of Medicine MU to junior researcher Z.K. (ROZV/28/LF/2020).
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Sumbal, J., Gudjonsson, T., Traustadottir, G.A., Koledova, Z. (2022). An Organotypic Assay to Study Epithelial-Fibroblast Interactions in Human Breast. In: Vivanco, M.d. (eds) Mammary Stem Cells. Methods in Molecular Biology, vol 2471. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2193-6_16
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2193-6_16
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2192-9
Online ISBN: 978-1-0716-2193-6
eBook Packages: Springer Protocols