Abstract
Background3D modelling fulfils a critical role in research, allowing for complex cell behaviour and interactions to be studied in physiomimetic conditions. With tissue banks becoming established for a number of cancers, researchers now have access to primary patient cells, providing the perfect building blocks to recreate and interrogate intricate cellular systems in the laboratory. The ducts of the human breast are composed of an inner layer of luminal cells supported by an outer layer of myoepithelial cells. In early-stage ductal carcinoma in situ, cancerous luminal cells are confined to the ductal space by an intact myoepithelial layer. Understanding the relationship between myoepithelial and luminal cells in the development of cancer is critical for the development of new therapies and prognostic markers. This requires the generation of new models that allows for the manipulation of these two cell types in a physiological setting.MethodsUsing access to the Breast Cancer Now Tissue Bank, we isolated pure populations of myoepithelial and luminal cells from human reduction mammoplasty specimens and placed them into 2D culture. These cells were infected with lentiviral particles encoding either fluorescent proteins, to facilitate cell tracking, or an inducible human epidermal growth factor receptor 2 (HER2) expression construct. Myoepithelial and luminal cells were then recombined in collagen gels, and the resulting cellular structures were analysed by confocal microscopy.Results Myoepithelial and luminal cells isolated from reduction mammoplasty specimens can be grown separately in 2D culture and retain their differentiated state. When recombined in collagen gels, these cells reform into physiologically reflective bilayer structures. Inducible expression of HER2 in the luminal compartment, once the bilayer has formed, leads to robust luminal filling, recapitulating ductal carcinoma in situ, and can be blocked with anti-HER2 therapies.ConclusionsThis model allows for the interaction between myoepithelial and luminal cells to be investigated in an in-vitro environment and paves the way to study early events in breast cancer development with the potential to act as a powerful drug discovery platform.
Highlights
3D modelling fulfils a critical role in research, allowing for complex cell behaviour and interactions to be studied in physiomimetic conditions
The ducts of the human breast are composed primarily of two cellular elements in a bilayer structure: luminal epithelial cells, which form a polarised layer around the central ductal cavity, and myoepithelial cells that are positioned between the basement membrane and the luminal epithelial layer
Primary human myoepithelial and luminal cells maintain their characteristics in vitro To adequately investigate the relationship between myoepithelial and luminal cells, these two cell populations first need to be separated and cultured individually to allow for their genetic manipulation prior to rebuilding the duct in vitro (Fig. 1a). These two cell types were separated from ductal organoids isolated from patients who had undergone reduction mammoplasty, based on their expression of CD10 and epithelial cell adhesion molecule (EpCAM), for myoepithelial and luminal cells, respectively (Fig. 1b)
Summary
Understanding the relationship between myoepithelial and luminal cells in the development of cancer is critical for the development of new therapies and prognostic markers This requires the generation of new models that allows for the manipulation of these two cell types in a physiological setting. The ducts of the human breast are composed primarily of two cellular elements in a bilayer structure: luminal epithelial cells, which form a polarised layer around the central ductal cavity, and myoepithelial cells that are positioned between the basement membrane and the luminal epithelial layer. These myoepithelial cells secrete extracellular matrix components required for the correct polarity. Combined with earlier detection of DCIS, there has been a rise in potential overdiagnosis of breast
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