动物造模 z-bio 2022-06-05 401

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

　　METHODS: The terminal ileum of 8-week-old C57BL/6 mice was collected and separated by EDTA method. The crypts were collected, embedded in Matrigel, and cultured in complete medium. Taking absolute alcohol and normal small intestine organoids as controls, short-term intervention with 100 μmol/L DCA at high concentration for 2 d, long-term intervention with 10 μmol/L DCA for 10 d, and after removing DCA, long-term culture for 10 d was performed to observe the growth of organoids and germination.

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

　　Conclusion: This study successfully established the mouse intestinal organoid culture technology. DCA damages the intestinal organoids and affects their growth, and their functions can be partially restored after long-term removal of DCA.