Molecular Profiling of Multiplexed Gene Markers to Assess Viability of Ex Vivo Human Colon Explant Cultures

Janice E Drew; Andrew J Farquharson; Hollie Vase; Frank A Carey; Robert J C Steele; Ruth A Ross; David C Bunton

doi:10.1089/biores.2015.0029

Molecular Profiling of Multiplexed Gene Markers to Assess Viability of Ex Vivo Human Colon Explant Cultures

Janice E Drew, Andrew J Farquharson, Hollie Vase, Frank A Carey, Robert J C Steele, Ruth A Ross, David C Bunton

The Rowett Institute of Nutrition and Health

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Abstract

Human colon tissue explant culture provides a physiologically relevant model system to study human gut biology. However, the small (20-30 mg) and complex tissue samples used present challenges for monitoring tissue stability, viability, and provision of sufficient tissue for analyses. Combining molecular profiling with explant culture has potential to overcome such limitations, permitting interrogation of complex gene regulation required to maintain gut mucosa in culture, monitor responses to culture environments and interventions. Human ex vivo colon explant gene expression profiles were assayed using an in-house custom-designed hCellMarkerPlex assay at culture time points 0, 1, 2, 4, and 14 h. Characteristic profiles of epithelial cell markers linked to differentiation, cellular polarization, and apoptosis were correlated with visible histochemical changes in explant epithelium during culture and tissue donors. The GenomeLab System provides effective assay of multiple targets not possible from small tissue samples with conventional gene expression technology platforms. This is advantageous to increase the utility of the ex vivo human colon model in applications to interrogate this complex and dynamic tissue environment for use in analytical testing.

Original language	English
Pages (from-to)	425-430
Number of pages	6
Journal	BioResearch Open Access
Volume	4
Issue number	1
DOIs	https://doi.org/10.1089/biores.2015.0029
Publication status	Published - 13 Nov 2015

Bibliographical note

© Janice E. Drew et al. 2015; Published by Mary Ann Liebert, Inc. This Open Access article is distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited.

Acknowledgments
The authors would like to thank the patients who kindly donated tissue samples, Sally Chalmers of the Tayside Tissue Bank for her help with collecting of the tissue donor samples, Emma Moss for advice on human colon dissection and explant culture, and Claus Dieter Mayer, Biomathematics and Statistics Scotland, for advice on statistical analysis. This work was supported by the Scottish Government (GT403), Scottish Universities Life Science Alliance, and TENOVUS Scotland.

Keywords

cell marker
gut explant
microanatomy
molecular profile
multiplex

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10.1089/biores.2015.0029Licence: CC BY

Molecular Profiling of Multiplexed Gene Markers to Assess Viability of Ex Vivo Human Colon Explant Cultures
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. http://creativecommons.org/licenses/by/4.0/
Final published version, 705 KBLicence: CC BY

Cite this

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title = "Molecular Profiling of Multiplexed Gene Markers to Assess Viability of Ex Vivo Human Colon Explant Cultures",

abstract = "Human colon tissue explant culture provides a physiologically relevant model system to study human gut biology. However, the small (20-30 mg) and complex tissue samples used present challenges for monitoring tissue stability, viability, and provision of sufficient tissue for analyses. Combining molecular profiling with explant culture has potential to overcome such limitations, permitting interrogation of complex gene regulation required to maintain gut mucosa in culture, monitor responses to culture environments and interventions. Human ex vivo colon explant gene expression profiles were assayed using an in-house custom-designed hCellMarkerPlex assay at culture time points 0, 1, 2, 4, and 14 h. Characteristic profiles of epithelial cell markers linked to differentiation, cellular polarization, and apoptosis were correlated with visible histochemical changes in explant epithelium during culture and tissue donors. The GenomeLab System provides effective assay of multiple targets not possible from small tissue samples with conventional gene expression technology platforms. This is advantageous to increase the utility of the ex vivo human colon model in applications to interrogate this complex and dynamic tissue environment for use in analytical testing.",

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Molecular Profiling of Multiplexed Gene Markers to Assess Viability of Ex Vivo Human Colon Explant Cultures

Abstract

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Keywords

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