Fig. 4.

Inflammatory induction in monolayer IECs by co-culturing with differentiated adipocytes. (a) Images of differentiated mouse embryonic fibroblasts (MEFs; days 0, 9, and 30) stained with Oil Red O. (b) Schematic of co-culture system for IECs and adipocytes. (c) Monolayer IECs derived from mouse small intestinal organoids were cultured for 13 days in Transwells, and then co-cultured in pre-stimulation medium for 2 days with MEFs (days 0, 7, and 26). IECs were then harvested, and the relative mRNA levels were determined by quantitative RT-PCR and normalized to GAPDH mRNA levels. (d) Images of differentiated human primary visceral adipocytes (days 0 and 14) stained with Oil Red O. (e) Monolayer IECs derived from human iPSC-derived organoids were cultured for 13 days in Transwells, and then co-cultured in pre-stimulation medium for 2 days with human primary adipocytes (days 0 and 11). IECs were then harvested, and the relative mRNA levels were determined by quantitative RT-PCR and normalized to GAPDH mRNA levels. The assay was performed in triplicate. Data are presented as mean ± SEM, *P < 0.05 (Student's t-test). All data are representative of at least two independent experiments.