Cargando…
Pre-Analytical Determination of the Effect of Extended Warm or Cold Ischaemia on RNA Stability in the Human Ileum Mucosa
The use of banked human tissue, obtained with informed consent after elective surgical procedures, represents a powerful model for understanding underlying mechanisms of diseases or therapeutic interventions and for establishing prognostic markers. However, donated tissues typically have varying tim...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4570714/ https://www.ncbi.nlm.nih.gov/pubmed/26371767 http://dx.doi.org/10.1371/journal.pone.0138214 |
_version_ | 1782390250479288320 |
---|---|
author | Lee, Serene M. L. Schelcher, Celine Thasler, Reinhard Schiergens, Tobias S. Thasler, Wolfgang E. |
author_facet | Lee, Serene M. L. Schelcher, Celine Thasler, Reinhard Schiergens, Tobias S. Thasler, Wolfgang E. |
author_sort | Lee, Serene M. L. |
collection | PubMed |
description | The use of banked human tissue, obtained with informed consent after elective surgical procedures, represents a powerful model for understanding underlying mechanisms of diseases or therapeutic interventions and for establishing prognostic markers. However, donated tissues typically have varying times of warm ischaemia in situ due to blood arrest or cold ischaemia due to procurement and transportation. Hence, before using these tissues, it is important to carry out pre-analytical studies to ensure that they are representative of the in vivo state. In particular, tissues of the gastrointestinal tract have been thought to have low RNA stability. Therefore, this study aimed to determine if extended warm or cold ischaemia times and snap-freezing or banking in RNA stabilization solution affects RNA integrity or gene expression in human ileum mucosa. In short, ileum mucosa was collected for up to 1.5 h and 6 h of simulated warm or cold ischaemia respectively. Subsequently, RNA integrity and gene expressions were determined. It was found that RNA integrity remained high over the course of warm and cold ischaemia examined and there were in general no significant differences between snap-freezing and banking in RNA stabilization solution. Following the same trend, there were in general no significant changes in gene expressions measured (MYC, HIF1α, CDX, HMOX1 and IL1β). In conclusion, RNA in the ileum mucosa is maintained at a high integrity and has stable gene expression over the examined time course of warm or cold ischaemia when banked in RNA stabilization solution or snap-frozen in liquid nitrogen. As the average warm and cold ischaemia times imposed by surgery and the process of tissue banking are shorter than the time period examined in this study, human ileum mucosa samples collected after surgeries could be used for gene expression studies. |
format | Online Article Text |
id | pubmed-4570714 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-45707142015-09-18 Pre-Analytical Determination of the Effect of Extended Warm or Cold Ischaemia on RNA Stability in the Human Ileum Mucosa Lee, Serene M. L. Schelcher, Celine Thasler, Reinhard Schiergens, Tobias S. Thasler, Wolfgang E. PLoS One Research Article The use of banked human tissue, obtained with informed consent after elective surgical procedures, represents a powerful model for understanding underlying mechanisms of diseases or therapeutic interventions and for establishing prognostic markers. However, donated tissues typically have varying times of warm ischaemia in situ due to blood arrest or cold ischaemia due to procurement and transportation. Hence, before using these tissues, it is important to carry out pre-analytical studies to ensure that they are representative of the in vivo state. In particular, tissues of the gastrointestinal tract have been thought to have low RNA stability. Therefore, this study aimed to determine if extended warm or cold ischaemia times and snap-freezing or banking in RNA stabilization solution affects RNA integrity or gene expression in human ileum mucosa. In short, ileum mucosa was collected for up to 1.5 h and 6 h of simulated warm or cold ischaemia respectively. Subsequently, RNA integrity and gene expressions were determined. It was found that RNA integrity remained high over the course of warm and cold ischaemia examined and there were in general no significant differences between snap-freezing and banking in RNA stabilization solution. Following the same trend, there were in general no significant changes in gene expressions measured (MYC, HIF1α, CDX, HMOX1 and IL1β). In conclusion, RNA in the ileum mucosa is maintained at a high integrity and has stable gene expression over the examined time course of warm or cold ischaemia when banked in RNA stabilization solution or snap-frozen in liquid nitrogen. As the average warm and cold ischaemia times imposed by surgery and the process of tissue banking are shorter than the time period examined in this study, human ileum mucosa samples collected after surgeries could be used for gene expression studies. Public Library of Science 2015-09-15 /pmc/articles/PMC4570714/ /pubmed/26371767 http://dx.doi.org/10.1371/journal.pone.0138214 Text en © 2015 Lee et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Lee, Serene M. L. Schelcher, Celine Thasler, Reinhard Schiergens, Tobias S. Thasler, Wolfgang E. Pre-Analytical Determination of the Effect of Extended Warm or Cold Ischaemia on RNA Stability in the Human Ileum Mucosa |
title | Pre-Analytical Determination of the Effect of Extended Warm or Cold Ischaemia on RNA Stability in the Human Ileum Mucosa |
title_full | Pre-Analytical Determination of the Effect of Extended Warm or Cold Ischaemia on RNA Stability in the Human Ileum Mucosa |
title_fullStr | Pre-Analytical Determination of the Effect of Extended Warm or Cold Ischaemia on RNA Stability in the Human Ileum Mucosa |
title_full_unstemmed | Pre-Analytical Determination of the Effect of Extended Warm or Cold Ischaemia on RNA Stability in the Human Ileum Mucosa |
title_short | Pre-Analytical Determination of the Effect of Extended Warm or Cold Ischaemia on RNA Stability in the Human Ileum Mucosa |
title_sort | pre-analytical determination of the effect of extended warm or cold ischaemia on rna stability in the human ileum mucosa |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4570714/ https://www.ncbi.nlm.nih.gov/pubmed/26371767 http://dx.doi.org/10.1371/journal.pone.0138214 |
work_keys_str_mv | AT leesereneml preanalyticaldeterminationoftheeffectofextendedwarmorcoldischaemiaonrnastabilityinthehumanileummucosa AT schelcherceline preanalyticaldeterminationoftheeffectofextendedwarmorcoldischaemiaonrnastabilityinthehumanileummucosa AT thaslerreinhard preanalyticaldeterminationoftheeffectofextendedwarmorcoldischaemiaonrnastabilityinthehumanileummucosa AT schiergenstobiass preanalyticaldeterminationoftheeffectofextendedwarmorcoldischaemiaonrnastabilityinthehumanileummucosa AT thaslerwolfgange preanalyticaldeterminationoftheeffectofextendedwarmorcoldischaemiaonrnastabilityinthehumanileummucosa |