Cryosectioning Method for Microdissection of Murine Colonic Mucosa

The colonic mucosal tissue provides a vital barrier to luminal antigens. This barrier is composed of a monolayer of simple columnar epithelial cells. The colonic epithelium is dynamically turned over and epithelial cells are generated in the stem cell containing crypts of Lieberkühn. Progenitor cell...

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Published inJournal of visualized experiments no. 101; p. e53112
Main Authors Farkas, Attila E, Gerner-Smidt, Christian, Lili, Loukia, Nusrat, Asma, Capaldo, Christopher T
Format Journal Article
LanguageEnglish
Published United States MyJove Corporation 12.07.2015
Subjects
Animals
Cell Differentiation - physiology
Cellular Biology
Colon - cytology
Cryoultramicrotomy - methods
Epithelial Cells - cytology
Fluorescent Antibody Technique - methods
Intestinal Mucosa - cytology
Mice
Mice, Inbred C57BL
Microdissection - methods
Microscopy, Confocal - methods
Stem Cells - cytology
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Summary:The colonic mucosal tissue provides a vital barrier to luminal antigens. This barrier is composed of a monolayer of simple columnar epithelial cells. The colonic epithelium is dynamically turned over and epithelial cells are generated in the stem cell containing crypts of Lieberkühn. Progenitor cells produced in the crypt-bases migrate toward the luminal surface, undergoing a process of cellular differentiation before being shed into the gut lumen. In order to study these processes at the molecular level, we have developed a simple method for the microdissection of two spatially distinct regions of the colonic mucosa; the proliferative crypt zone, and the differentiated surface epithelial cells. Our objective is to isolate specific crypt and surface epithelial cell populations from mouse colonic mucosa for the isolation of RNA and protein.
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Correspondence to: Christopher T. Capaldo at ccapald@emory.edu
ISSN:1940-087X
1940-087X
DOI:10.3791/53112