Comparison of four primary culture methods for mouse intestinal epithelial cells

【Objective】 Separation results of four primary culture methods for mouse intestinal epithelial cells were compared to choose and establish utility separation culture method for obtaining mouse IECs and preparing for future research.【Method】 The four methods,tissue fractional cultivation,thermolysin enzyme digestion,association digestion of collagenase XI and dispase I,and association digestion collagenase I and dispaseⅥ were used to separte and culture mouse IECs.The separation results were compared by the cell immunohistochemistry method and cell immunofluorescence method to identify the cell specificity of the separated IECs.【Result】 The results showed that the intestinal epithelial cells obtained by tissue culture method had the highest proliferation ability and better viability,but with serious fibroblasts pollution and lower purity.The collagenase I and neutral proteaseⅥ method only obtained a few intestinal epithelial cells with lower proliferation.The rmolysin digestion method and the joint digestion method of collagenase XI and neutral protease I obtained intestinal epithelial cells with higher purity and the proliferation ability was slightly weaker than the tissue culture method.However,the proliferation ability was more stabilized.The results of cell specificity of the separated IECs by the two methods using the cell immunohistochemistry method and cell immunofluorescence method showed that most of the separated cells were IECs.【Conclusion】 Thermolysin digestion method and Collagenase Ⅺ and Dispase Ⅰ combined digestion method are suitable for isolating and cultivating Intestinal epithelial cells.