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Differentiation of Human Colon Tissue In Culture: Effects of calcium on trans-epithelial electrical resistance and tissue cohesive properties

Shannon D McClintock, View ORCID ProfileMichael K Dame, Aliah Richter, Durga Attili, Sabrina S Silvestri, View ORCID ProfileMaliha M Berner, View ORCID ProfileMargaret S Bohm, Kateryna Karpoff, Caroline McCarthy, Jason R Spence, James Varani, View ORCID ProfileMuhammad N Aslam
doi: https://doi.org/10.1101/744979
Shannon D McClintock
1Department of Pathology, The University of Michigan Medical School, Ann Arbor, Michigan 48109, United States of America
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Michael K Dame
2Department of Internal Medicine (The Division of Gastroenterology), The University of Michigan Medical School, Ann Arbor, Michigan 48109, United States of America
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Aliah Richter
1Department of Pathology, The University of Michigan Medical School, Ann Arbor, Michigan 48109, United States of America
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Durga Attili
1Department of Pathology, The University of Michigan Medical School, Ann Arbor, Michigan 48109, United States of America
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Sabrina S Silvestri
2Department of Internal Medicine (The Division of Gastroenterology), The University of Michigan Medical School, Ann Arbor, Michigan 48109, United States of America
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Maliha M Berner
2Department of Internal Medicine (The Division of Gastroenterology), The University of Michigan Medical School, Ann Arbor, Michigan 48109, United States of America
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Margaret S Bohm
2Department of Internal Medicine (The Division of Gastroenterology), The University of Michigan Medical School, Ann Arbor, Michigan 48109, United States of America
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Kateryna Karpoff
2Department of Internal Medicine (The Division of Gastroenterology), The University of Michigan Medical School, Ann Arbor, Michigan 48109, United States of America
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Caroline McCarthy
2Department of Internal Medicine (The Division of Gastroenterology), The University of Michigan Medical School, Ann Arbor, Michigan 48109, United States of America
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Jason R Spence
2Department of Internal Medicine (The Division of Gastroenterology), The University of Michigan Medical School, Ann Arbor, Michigan 48109, United States of America
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James Varani
1Department of Pathology, The University of Michigan Medical School, Ann Arbor, Michigan 48109, United States of America
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Muhammad N Aslam
1Department of Pathology, The University of Michigan Medical School, Ann Arbor, Michigan 48109, United States of America
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  • For correspondence: mnaslam@med.umich.edu
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Abstract

Background and aims 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 minerals), there is little change in tight junction proteins but a strong up-regulation of desmosomal proteins and an increase in desmosome formation. The aim of this study was to assess functional consequences of the differences in calcium-mediated barrier protein expression.

Methods Human colonoid-derived epithelial monolayers were interrogated in transwell culture under low- or high-calcium conditions. Ion permeability and monolayer integrity were assessed by measuring trans-epithelial electrical resistance (TEER) across the confluent monolayer. Colonoid cohesiveness was assessed in parallel.

Results TEER values were high in the low-calcium environment and increased only modestly in response to calcium. In contrast, colonoid cohesiveness increased substantially with calcium supplementation. In both assays, the response to Aquamin was greater than the response to calcium alone. However, differences between interventions were small and only compared to the 0.25 mM calcium were they statistically significant. Consistent with these findings, occludin expression (a measure of tight junctions) was high at 0.25 mM calcium and did not increase with supplementation. Cadherin-17 and desmoglein-2 were weakly-expressed under low calcium conditions but increased with intervention.

Conclusions These findings indicate that low ambient calcium levels are sufficient to support formation of a permeability barrier in the colonic epithelium. Higher calcium levels are necessary to promote tissue cohesion and enhance barrier function. These findings may help explain how an adequate daily intake of calcium contributes to colonic health by improving barrier function, even though there is little change in colonic histological features over a wide range of calcium intake levels.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted August 22, 2019.
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Differentiation of Human Colon Tissue In Culture: Effects of calcium on trans-epithelial electrical resistance and tissue cohesive properties
Shannon D McClintock, Michael K Dame, Aliah Richter, Durga Attili, Sabrina S Silvestri, Maliha M Berner, Margaret S Bohm, Kateryna Karpoff, Caroline McCarthy, Jason R Spence, James Varani, Muhammad N Aslam
bioRxiv 744979; doi: https://doi.org/10.1101/744979
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Differentiation of Human Colon Tissue In Culture: Effects of calcium on trans-epithelial electrical resistance and tissue cohesive properties
Shannon D McClintock, Michael K Dame, Aliah Richter, Durga Attili, Sabrina S Silvestri, Maliha M Berner, Margaret S Bohm, Kateryna Karpoff, Caroline McCarthy, Jason R Spence, James Varani, Muhammad N Aslam
bioRxiv 744979; doi: https://doi.org/10.1101/744979

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