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A low-temperature method for the isolation of small-intestinal epithelium along the crypt-villus axis.

dc.contributor.authorFlint, Neil
dc.contributor.authorCove, Frances L
dc.contributor.authorEvans, Gareth S
dc.date.accessioned2010-06-21T14:04:49Z
dc.date.available2010-06-21T14:04:49Z
dc.date.issued1991-12-01
dc.identifier.citationA low-temperature method for the isolation of small-intestinal epithelium along the crypt-villus axis. 1991, 280 ( Pt 2):331-4 Biochem. J.en
dc.identifier.issn0264-6021
dc.identifier.pmid1747105
dc.identifier.urihttp://hdl.handle.net/10541/106582
dc.description.abstractA variety of enzymic and non-enzymic methods to isolate epithelium from the small intestine have been previously published. Sequential fractionation of cells from the villus to the crypt has been reported in some of these papers, which allows the comparative study of terminally differentiated and proliferative cell phenotypes. However, these methods often involve the incubation of tissues at 37 degrees C, which may affect the structural and biochemical integrity of the cells. We have developed a rapid low-temperature (4 degrees C) method for isolating purified populations of crypt and villus cells from mouse and rat intestines. The fractionated cells have been partially characterized, and the potential value of the procedure has been indicated by the ability to analyse the comparative protein and mRNA expression along the crypt-villus axis.
dc.language.isoenen
dc.subject.meshAlkaline Phosphatase
dc.subject.meshAnimals
dc.subject.meshCell Fractionation
dc.subject.meshCells, Cultured
dc.subject.meshCold Temperature
dc.subject.meshElectrophoresis, Agar Gel
dc.subject.meshEpithelial Cells
dc.subject.meshEpithelium
dc.subject.meshIntestine, Small
dc.subject.meshMale
dc.subject.meshProteins
dc.subject.meshRNA, Messenger
dc.subject.meshRats
dc.subject.meshRats, Inbred Strains
dc.subject.meshThymidine
dc.titleA low-temperature method for the isolation of small-intestinal epithelium along the crypt-villus axis.en
dc.typeArticleen
dc.contributor.departmentDepartment of Epithelial Biology, Paterson Institute for Cancer Research, Christie Hospital, Withington, Manchester, U.K.en
dc.identifier.journalBiochemical Journalen
html.description.abstractA variety of enzymic and non-enzymic methods to isolate epithelium from the small intestine have been previously published. Sequential fractionation of cells from the villus to the crypt has been reported in some of these papers, which allows the comparative study of terminally differentiated and proliferative cell phenotypes. However, these methods often involve the incubation of tissues at 37 degrees C, which may affect the structural and biochemical integrity of the cells. We have developed a rapid low-temperature (4 degrees C) method for isolating purified populations of crypt and villus cells from mouse and rat intestines. The fractionated cells have been partially characterized, and the potential value of the procedure has been indicated by the ability to analyse the comparative protein and mRNA expression along the crypt-villus axis.


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