Adult Stem Cell-Derived Intestinal Organoids as In Vitro Models of High-Fat Diet-Related Intestinal Diseases.

The dysregulation of intestinal lipid metabolism induced by a high-fat diet (HFD) is associated with metabolic diseases; however, the validity of intestinal organoids (IOs) as substitutes for traditional research systems remains uncertain. This study aimed to determine whether fatty acid (FA)-treated IOs accurately replicate intestinal lipid metabolism observed in HFD mice. Male C57BL/6 mice were fed either a normal chow diet (NCD) or HFD for 12 weeks, while mouse small intestinal crypt-derived IOs were treated in vitro with either an FA cocktail or a vehicle. Mice on the HFD exhibited phenotypes characteristic of metabolic syndrome, including intestinal lipid accumulation, upregulation of lipid catabolic genes, and downregulation of lipogenic genes. FA-treated IOs demonstrated enhanced budding frequency, lipid droplet accumulation, enriched lipid catabolism pathways, and suppressed lipogenesis, aligning with the in vivo findings. Omics analyses of the top 87 variable genes indicated a strong congruence between FA-treated IOs and intestinal tissues from HFD mice, with control groups clustering distinctly. The differentially expressed genes in both models were implicated in metabolic reprogramming, immune modulation, and barrier remodeling. Therefore, FA-treated IOs recapitulate key morphological and transcriptional characteristics of intestinal lipid metabolism in HFD mice, offering a valuable complementary model for investigating intestinal metabolic disorders.