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The development of a high throughput drug-responsive model of white adipose tissue comprising adipogenic 3T3-L1 cells in a 3D matrix.

Biofabrication (2019-11-13)
Alexander D Graham, Rajesh Pandey, Viktoriya S Tsancheva, Alessia Candeo, Stanley W Botchway, Alasdair J Allan, Lydia Teboul, Kamel Madi, Tahkur S Babra, Louisa A K Zolkiewski, Xuan Xue, Liz Bentley, Joan Gannon, Sam N Olof, Roger D Cox
ABSTRACT

Adipose models have been applied to mechanistic studies of metabolic diseases (such as diabetes) and the subsequent discovery of new therapeutics. However, typical models are either insufficiently complex (2D cell cultures) or expensive and labor intensive (mice/in vivo). To bridge the gap between these models and in order to better inform pre-clinical studies we have developed a drug-responsive 3D model of white adipose tissue (WAT). Here, spheroids (680 ± 60 μm) comprising adipogenic 3T3-L1 cells encapsulated in 3D matrix were fabricated manually on a 96 well scale. Spheroids were highly characterised for lipid morphology, selected metabolite and adipokine secretion, and gene expression; displaying significant upregulation of certain adipogenic-specific genes compared with a 2D model. Furthermore, induction of lipolysis and promotion of lipogenesis in spheroids could be triggered by exposure to 8-br-cAMP and oleic-acid respectively. Metabolic and high content imaging data of spheroids exposed to an adipose-targeting drug, rosiglitazone, resulted in dose-responsive behavior. Thus, our 3D WAT model has potential as a powerful scalable tool for compound screening and for investigating adipose biology.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Propidium iodide, ≥94.0% (HPLC)
Sigma-Aldrich
Forskolin, For use in molecular biology applications
Sigma-Aldrich
Oleic acid, suitable for cell culture, BioReagent
Sigma-Aldrich
Rosiglitazone, ≥98% (HPLC)
Sigma-Aldrich
Mineral oil, suitable for preparation of Nujol mulls for infrared spectroscopy, light oil
Sigma-Aldrich
8-Bromoadenosine 3′,5′-cyclic monophosphate sodium salt, ≥97% (HPLC), powder
Sigma-Aldrich
Hexadecane, anhydrous, ≥99%
Sigma-Aldrich
Undecane, ≥99%
Sigma-Aldrich
Silicone oil AR 20, viscosity ~20 mPa.s,   neat(25 °C)
Sigma-Aldrich
Adenosine 3′,5′-cyclic monophosphate tris salt, ≥97% (HPLC), powder