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A Primary Cell and Organoid Platform for Evaluating Pharmacological Responses in Mammary Epithelial Cells.

ACS pharmacology & translational science (2020-04-08)
Teneale A Stewart, Felicity M Davis
ABSTRACT

An essential process in predicting the in vivo pharmacological activity of a candidate molecule involves the evaluation of target responses using established model systems. While these models largely comprise immortalized cells, which are often serially passaged as monolayers on uniformly stiff substrates and are modified to overexpress one or more components of the pathway-of-interest, the importance of cell identity, heterogeneity, and three-dimensional (3D) context to target response is gaining increasing attention. Here, we assess intracellular calcium responses in mouse mammary epithelial cells in three distinct model systems: 3D primary organoids, 2D primary epithelial cells, and 2D immortalized cells. Specifically, we assess intracellular calcium responses to a number of extracellular signals implicated in the regulation of basal (or myoepithelial) cell function. These findings provide further insights into cell type and context-specific pharmacological responses in mammary epithelial cells and highlight the opportunities and challenges in the adoption of architecturally complex and heterogeneous in vitro assays in pharmacological research.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
RPMI-1640 Medium, With L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Insulin from bovine pancreas, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Adenosine 5′-triphosphate disodium salt hydrate, microbial, BioReagent, suitable for cell culture, ≥99% (HPLC)
Sigma-Aldrich
Sunflower seed oil from Helianthus annuus
Sigma-Aldrich
Epidermal Growth Factor from murine submaxillary gland, EGF, suitable for cell culture
Sigma-Aldrich
Oxytocin