Analysis of the Biochemical Integrity of the Isolated Mouse Small Intestinal Crypt and Villus Cells

Objective: To test the biochemical integrity of mouse intestinal crypt and villus cells isolated and enriched at low temperature using chelating agents. Methods: The crypt and villus units of small intestinal epithelium were isolated and enriched at low temperature (ice cold) using chelating agents. The genome DNA, total RNA and total protein were extracted and electrophoresed. The mR- NA expression level of lysozyme was detected by real-time PCR to evaluate the purity of enriched crypt and villus units. Results: The enriched crypt and villus units remained morphological integrity. The integrity of genome DNA, total RNA and protein were preserved during isolation procedure. The expression level of lysozyme in the enriched crypt units is more than 30 times higher than that in villus units. Conclusion: The structural and biochemical integrity of crypt and villus cells can be preserved using the improved low-temperature method. In addition, this method may be useful for in vivo analysis of protein and mRNA expression in normal and stressed small intestinal epithelial cells.