Differentiation of human colon tissue in culture: Effects of calcium on trans-epithelial electrical resistance and tissue cohesive properties.

PloS One
Shannon D McClintockMuhammad N Aslam

Abstract

Human colonoid cultures maintained under low-calcium (0.25 mM) conditions undergo differentiation spontaneously and, concomitantly, express a high level of tight junction proteins, but not desmosomal proteins. When calcium is included to a final concentration of 1.5-3.0 mM (provided either as a single agent or as a combination of calcium and additional minerals), there is little change in tight junction protein expression but a strong up-regulation of desmosomal proteins and an increase in desmosome formation. The aim of this study was to assess the functional consequences of calcium-mediated differences in barrier protein expression. Human colonoid-derived epithelial cells were interrogated in transwell culture under low- or high-calcium conditions for monolayer integrity and ion permeability by measuring trans-epithelial electrical resistance (TEER) across the confluent monolayer. Colonoid cohesiveness was assessed in parallel. TEER values were high in the low-calcium environment but increased in response to calcium. In addition, colonoid cohesiveness increased substantially with calcium supplementation. In both assays, the response to multi-mineral intervention was greater than the response to calcium alone. Consistent with ...Continue Reading

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Citations

Dec 18, 2020·Frontiers in Cell and Developmental Biology·Muhammad N AslamJames Varani
Mar 25, 2021·American Journal of Physiology. Gastrointestinal and Liver Physiology·Paula Marincola SmithR Daniel Beauchamp
Oct 30, 2021·Antioxidants & Redox Signaling·Aleksandr VendrovWilliam J Arendshorst

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Methods Mentioned

BETA
transmission electron microscopy
biopsies
confocal fluorescence microscopy
fluorescence microscopy
electron microscopy

Clinical Trials Mentioned

NCT02647671

Software Mentioned

Aquamin
GraphPad Prism
Adobe Photoshop
Formats Importer
Bio
ImageJ
Fiji

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