Three-Dimensional Culture of Human Breast Epithelial Cells: The How and the Why

Abstract

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.

Culture phases, cytotoxicity and protein expressions of agarose hydrogel induced Sp2/0, A549, MCF-7 cell line 3D cultures

“…The trend in terms of increasing cell culture complexity is towards 3 dimensional (3D) cultures that make it possible to create ex vivo conditions in the lab. 3D cell cultures have proven to be very useful for several studies including cell physiology, cell behaviour, cellular metabolism, cytotoxicity, genotoxicity, biomarker discovery, cell development and differentiation, protein and gene expression and tissue engineering applications (Pampaloni et al 2007;Longati et al 2013;Vidi et al 2013).…”

3D Miniaturization of Human Organs for Drug Discovery

“…[81,[168][169][170][171] Various microfluidic models have been developed for studying tumor angiogenesis, [81] intravasation, [170] the role of interstitial flow, [171] and matrix stiffness [172] on cancer cell migration with adhesion, [173] and extravasation. [168] Effects of interstitial flow on tumor migration were examined to better understand how cancer cells extravasate from the blood vessels.…”

“…[140] In these studies, since basal cells were not cloned post-transfection, the used cell populations carried a mixture of mutant GRHL2 alleles. While conditions have been developed in which single basal cells can be cloned in 2D culture, [169] whether post-expansion each clone can form organoids remains to be tested. Additionally, nasospheres, organoids derived from nasal cells, potentially could allow the screening of small molecules and drugs that may regulate or compensate for the activity of mutant forms of the cystic fibrosis transmembrane conductance regulator, as measured by fluid transport and sphere diameter.…”

Quantification of Breast Cancer Cell Invasiveness Using a Three-dimensional (3D) Model

“…The maintenance of breast tissue integrity depends on tightly controlled tissue architecture since cell-ECM and cell-cell adhesion junctions influence gene expression and disruption of epithelial polarity can lead to the onset of cancer [6][7][8][9][10] . However, most in vitro migration and invasion assays such as transwell chamber assays or wound-scratch assays are two-dimensional (2D) and hence these neglect the intricate interactions between cells and their adjacent environment 3,6,8,[11][12][13][14] .…”