Three-Dimensional Culture of Human Breast Epithelial Cells: The How and the Why
Organs are made of the organized assembly of different cell types that contribute to the architecture necessary for functional differentiation. In those with exocrine function, such as the breast, cell–cell and cell–extracellular matrix (ECM) interactions establish mechanistic constraints and a complex biochemical signaling network essential for differentiation and homeostasis of the glandular epithelium. Such knowledge has been elegantly acquired for the mammary gland by placing epithelial cells under three-dimensional (3D) culture conditions.
Three-dimensional cell culture aims at recapitulating normal and pathological tissue architectures, hence providing physiologically relevant models to study normal development and disease. The specific architecture of the breast epithelium consists of glandular structures (acini) connected to a branched ductal system. A single layer of basoapically polarized luminal cells delineates ductal or acinar lumena at the apical pole. Luminal cells make contact with myoepithelial cells and, in certain areas at the basal pole, also with basement membrane (BM) components. In this chapter, we describe how this exquisite organization as well as stages of disorganization pertaining to cancer progression can be reproduced in 3D cultures. Advantages and limitations of different culture settings are discussed. Technical designs for induction of phenotypic modulations, biochemical analyses, and state-of-the-art imaging are presented. We also explain how signaling is regulated differently in 3D cultures compared to traditional two-dimensional (2D) cultures. We believe that using 3D cultures is an indispensable method to unravel the intricacies of human mammary functions and would best serve the fight against breast cancer.
Key words3D culture Breast epithelium Basoapical polarity Cancer progression Tissue architecture
We thank Dr. Kurt Hodges for providing micrographs from tissue sections used in Fig. 1. Support was provided by the National Institutes of Health (R01CA112017 and R03CA112613) and the Susan G. Komen Breast Cancer Foundation (BCTR-0707641) to SAL; the US Department of Energy, Office of Biological and Environmental Research, via a Distinguished Fellow Award and Low Dose Radiation Program (DE-AC02-05CH1123), the National Institutes of Health (R37CA064786, U54CA126552, R01CA057621, U54CA112970, U01CA143233, U54CA143836—Bay Area Physical Sciences–Oncology Center, University of California, Berkeley, California), and the US Department of Defense (W81XWH0810736) to MJB; postdoctoral fellowships from the Novartis Foundation and the Swiss National Science Foundation (PBNEA–116967) to PAV.
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