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

Biochemical Journal - Tập 280 Số 2 - Trang 331-334 - 1991
Neil Flint1, FRANCES L. COVE1, Gareth Evans1
1Cancer Research Campaign, Department of Epithelial Biology, Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute, Wilmslow Road, Withington, Manchester M20 9BX, U.K.

Tóm tắt

A 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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