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  • Scaffold architecture controls insulinoma clustering, viability, and insulin production.

Scaffold architecture controls insulinoma clustering, viability, and insulin production.

Tissue engineering. Part A (2014-01-15)
Britani N Blackstone, Andre F Palmer, Horacio R Rilo, Heather M Powell
ABSTRACT

Recently, in vitro diagnostic tools have shifted focus toward personalized medicine by incorporating patient cells into traditional test beds. These cell-based platforms commonly utilize two-dimensional substrates that lack the ability to support three-dimensional cell structures seen in vivo. As monolayer cell cultures have previously been shown to function differently than cells in vivo, the results of such in vitro tests may not accurately reflect cell response in vivo. It is therefore of interest to determine the relationships between substrate architecture, cell structure, and cell function in 3D cell-based platforms. To investigate the effect of substrate architecture on insulinoma organization and function, insulinomas were seeded onto 2D gelatin substrates and 3D fibrous gelatin scaffolds with three distinct fiber diameters and fiber densities. Cell viability and clustering was assessed at culture days 3, 5, and 7 with baseline insulin secretion and glucose-stimulated insulin production measured at day 7. Small, closely spaced gelatin fibers promoted the formation of large, rounded insulinoma clusters, whereas monolayer organization and large fibers prevented cell clustering and reduced glucose-stimulated insulin production. Taken together, these data show that scaffold properties can be used to control the organization and function of insulin-producing cells and may be useful as a 3D test bed for diabetes drug development.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
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Sodium pyruvate, BioXtra, ≥99%
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Sodium pyruvate, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99%
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Sodium pyruvate, ReagentPlus®, ≥99%
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2-Mercaptoethanol, Molecular Biology, suitable for electrophoresis, suitable for cell culture, BioReagent, 99% (GC/titration)
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2-Mercaptoethanol, ≥99.0%
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2-Mercaptoethanol, BioUltra, Molecular Biology, ≥99.0% (GC)
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Methane-12C, 13C-depleted, 99.9 atom % 12C
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N,O-Bis(trimethylsilyl)acetamide, synthesis grade, ≥95%
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HEPES buffer solution, 1 M in H2O
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DAPI, for nucleic acid staining