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 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.
Epithelial Pathobiology and Mucosal Inflammation Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia.
Attila E
AE
Farkas
Epithelial Pathobiology and Mucosal Inflammation Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia.
Christian
C
Gerner-Smidt
Epithelial Pathobiology and Mucosal Inflammation Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia.
Loukia
L
Lili
Epithelial Pathobiology and Mucosal Inflammation Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia.
Asma
A
Nusrat
Epithelial Pathobiology and Mucosal Inflammation Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia; ccapald@emory.edu.
Christopher T
CT
Capaldo
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D003106:Colon / Q000166:cytology*; D016616:Cryoultramicrotomy / Q000379:methods*; D004847:Epithelial Cells / Q000166:cytology; D005455:Fluorescent Antibody Technique / Q000379:methods; D007413:Intestinal Mucosa / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D042282:Microdissection / Q000379:methods*; D018613:Microscopy, Confocal / Q000379:methods; D013234:Stem Cells / Q000166:cytology