Separation of intact intestinal epithelium from mesenchyme

Current protocols for separating adult intestinal epithelial cells from the underlying muscular and mesenchymal tissues typically involve extended incubations, harsh mechanical treatment, and exposure to either proteases or chelating agents. The drawbacks of these approaches include fragmentation, c...

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Main Authors: Ali Moussavi Nik, Peter Carlsson
Format: Article
Language:English
Published: Taylor & Francis Group 2013-07-01
Series:BioTechniques
Subjects:
Online Access:https://www.future-science.com/doi/10.2144/000114055
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author Ali Moussavi Nik
Peter Carlsson
author_facet Ali Moussavi Nik
Peter Carlsson
author_sort Ali Moussavi Nik
collection DOAJ
description Current protocols for separating adult intestinal epithelial cells from the underlying muscular and mesenchymal tissues typically involve extended incubations, harsh mechanical treatment, and exposure to either proteases or chelating agents. The drawbacks of these approaches include fragmentation, contamination with other cell types, reduced viability, and under-representation of crypt cells. Here we describe a gentle procedure that allows harvesting of pure, fully viable sheets of murine intestinal epithelium, with intact crypts and villi, without enzymes or EDTA. The mesenchyme retains intact villus core projections, is virtually free from epithelial cells, and can be cultured in vitro.
format Article
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1940-9818
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spelling doaj-art-0d582153a6a04561b337cb31cb4a92502025-08-20T02:26:03ZengTaylor & Francis GroupBioTechniques0736-62051940-98182013-07-01551424410.2144/000114055Separation of intact intestinal epithelium from mesenchymeAli Moussavi Nik0Peter Carlsson11Department of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, Sweden1Department of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, SwedenCurrent protocols for separating adult intestinal epithelial cells from the underlying muscular and mesenchymal tissues typically involve extended incubations, harsh mechanical treatment, and exposure to either proteases or chelating agents. The drawbacks of these approaches include fragmentation, contamination with other cell types, reduced viability, and under-representation of crypt cells. Here we describe a gentle procedure that allows harvesting of pure, fully viable sheets of murine intestinal epithelium, with intact crypts and villi, without enzymes or EDTA. The mesenchyme retains intact villus core projections, is virtually free from epithelial cells, and can be cultured in vitro.https://www.future-science.com/doi/10.2144/000114055intestineepitheliummesenchymeseparation
spellingShingle Ali Moussavi Nik
Peter Carlsson
Separation of intact intestinal epithelium from mesenchyme
BioTechniques
intestine
epithelium
mesenchyme
separation
title Separation of intact intestinal epithelium from mesenchyme
title_full Separation of intact intestinal epithelium from mesenchyme
title_fullStr Separation of intact intestinal epithelium from mesenchyme
title_full_unstemmed Separation of intact intestinal epithelium from mesenchyme
title_short Separation of intact intestinal epithelium from mesenchyme
title_sort separation of intact intestinal epithelium from mesenchyme
topic intestine
epithelium
mesenchyme
separation
url https://www.future-science.com/doi/10.2144/000114055
work_keys_str_mv AT alimoussavinik separationofintactintestinalepitheliumfrommesenchyme
AT petercarlsson separationofintactintestinalepitheliumfrommesenchyme