A low-temperature method for the isolation of small-intestinal epithelium along the crypt-villus axis.
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.
Flint
N
N
Department of Epithelial Biology, Paterson Institute for Cancer Research, Christie Hospital, Withington, Manchester, U.K.
Cove
F L
FL
Evans
G S
GS
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002478:Cells, Cultured; D003080:Cold Temperature; D004587:Electrophoresis, Agar Gel; D004847:Epithelial Cells; D004848:Epithelium / Q000201:enzymology / Q000378:metabolism; D007421:Intestine, Small / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism; D008297:Male; D011506:Proteins / Q000378:metabolism; D012333:RNA, Messenger / Q000302:isolation & purification / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013936:Thymidine / Q000378:metabolism
D011506:Proteins; D012333:RNA, Messenger; D000469:Alkaline Phosphatase; D013936:Thymidine