A 3D Fibroblast-Epithelium Co-culture Model for Understanding Microenvironmental Role in Branching Morphogenesis of the Mammary Gland

  • Zuzana Koledova
  • Pengfei Lu
Part of the Methods in Molecular Biology book series (MIMB, volume 1501)


The mammary gland consists of numerous tissue compartments, including mammary epithelium, an array of stromal cells, and the extracellular matrix (ECM). Bidirectional interactions between the epithelium and its surrounding stroma are essential for proper mammary gland development and homeostasis, whereas their deregulation leads to developmental abnormalities and cancer. To study the relationships between the epithelium and the stroma, development of models that could recapitulate essential aspects of these interacting systems in vitro has become necessary. Here we describe a three-dimensional (3D) co-culture assay and show that the addition of fibroblasts to mammary organoid cultures promotes the epithelium to undergo branching morphogenesis, thus allowing the role of the stromal microenvironment to be examined in this essential developmental process.

Key words

3D culture Branching morphogenesis Extracellular matrix Fibroblasts Mammary epithelium Matrigel Organoids Organotypic assay Paracrine signaling 



This work was supported by grants from the National Institutes of Health (R03 HD060807), Breast Cancer Now, and ShanghaiTech University (to P.L.), and by the project “Employment of Best Young Scientists for International Cooperation Empowerment“(CZ.1.07/2.3.00/30.0037) co-financed from European Social Fund and the state budget of the Czech Republic and by the grant “Junior investigator 2015“ from Faculty of Medicine, Masaryk University (to Z.K.). The Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, was supported by core funding from the Wellcome Trust (088785/Z/09/Z).


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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Wellcome Trust Centre for Cell Matrix Research, Faculty of Life SciencesUniversity of ManchesterManchesterUK
  2. 2.Department of Histology and Embryology, Faculty of MedicineMasaryk UniversityBrnoCzech Republic
  3. 3.Department of Anatomy and Program in Developmental and Stem Cell BiologyUniversity of CaliforniaSan FranciscoUSA
  4. 4.School of Life Science and TechnologyShanghai Tech UniversityShanghaiChina

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