动物造模 z-bio 2021-09-27 275

　　Objective: To establish a mouse small intestinal organoid culture system and discuss the effect of deoxycholic acid (DCA) on the growth of small intestinal organoids.

　　Method: Take 8 weeks old C57BL/6 mouse terminal ileum, separate by EDTA method, collect crypts, embed in Matrigel matrigel, and cultivate in intact culture medium. Take anhydrous alcohol and normal small intestine organoids as controls, give 100 μmol/L high-concentration DCA short-term intervention for 2 d, 10 μmol/L DCA long-term intervention for 10 d, and remove DCA and incubate for a long-term 10 d to observe the growth of organoids And budding status.

　　Results: Short-term intervention of high-concentration DCA in organoids, the composition rate of intestinal organoid spherical structure, intestinal organoid structure [composition rate, organoid evolution rate and number of budding were significantly reduced (P<0.05), short-term removal of the four indicators of DCA It still decreased (P<0.05). After long-term removal of DCA, only the intestinal organoid structure and the number of buds decreased (P<0.05). Low-concentration DCA short-term intervention of intestinal organoid structure and the number of buds decreased (P<0.05), long-term intervention of intestinal organoid spherical structure, the rate of intestinal organoid structure and the number of buds were significantly reduced (P<0.05) ), after removing DCA, only intestinal organoid structure composition rate and number of buds were lower than normal (P<0.05).

　　Conclusion: This study successfully established the mouse small intestinal organoid culture technology. DCA damages the small intestinal organoid and affects its growth. After long-term removal of DCA, its function can be partially restored.