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Separation of intact intestinal epithelium from mesenchyme

Journal article
Authors Ali Moussavi Nik
Peter Carlsson
Published in Biotechniques
Volume 55
Issue 1
Pages 42-44
ISSN 0736-6205
Publication year 2013
Published at Department of Chemistry and Molecular Biology
Pages 42-44
Language en
Keywords intestine, epithelium, mesenchyme, separation, pig small-intestine, endocrine-cells, stem-cells, mouse
Subject categories Medical Biotechnology, Biochemistry and Molecular Biology


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..

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