Fig. 4: Organoid-derived MA controls macrophage polarization.

A Schematic illustration of the experimental set-up. B Concentration of IL-10 in the supernatant of BMDMs co-cultured with organoid-derived conditioned or control medium determined by ELISA (n = 5 biological replicates for control group, n = 6 biological replicates for CM group). Represent significant ****p value <0.0001 using two-tailed student’s t test. C Quantification of CD206⁺F4/80⁺CD11b⁺ macrophages and CD206 MFI (n = 4 biological replicates for control group, n = 5 biological replicates for CM group). CD206⁺F4/80⁺CD11b⁺ macrophages: represent significant *p value = 0.0462 using two-tailed student’s t test; CD206 MFI: represent significant *p value = 0.0389 using two-tailed student’s t test. D Representative images of immunostaining of CD206 (green) and DAPI (blue) in BMDMs stimulated with organoid-derived conditioned or control medium. E Genetic profiling of BMDMs stimulated with organoid-derived conditioned medium or control medium (n = 6 biological replicates/group). Represent significant p value using two-tailed student’s from left to right: 0.0037, 0.0036, 0.0451. F Top 35 metabolites in the organoid-derived conditioned medium group compared with those in the control group under negative ion modes using untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomic analysis. G Volcano plot displaying the different metabolites in the cecal contents of organoid-transplanted and control mice under negative ion modes after I/R using untargeted LC-MS metabolomic analysis. Venn diagram displaying the consensus upregulated metabolites in the organoid-derived conditioned medium group and cecal contents of organoid-transplanted mice compared with those in control mice under positive (H) and negative (I) ion modes, respectively. J MA levels in organoid-derived conditioned and control medium groups using targeted LC-MS metabolomic analysis (n = 3 biological replicates for control group, n = 5 biological replicates for CM group). Represent significant **p value = 0.0047 using two-tailed student’s t test. K MA levels in the cecal contents of the organoid-transplanted and control groups after I/R using targeted LC-MS metabolomic analysis (n = 7 mice for control group, n = 9 mice for transplanted group). Represent significant **p value = 0.0012 using two-tailed Mann–Whitney test. L Correlation analysis between preoperative patient fecal MA content and LIFE score (n = 23 samples/group). M Correlation analysis between preoperative fecal MA levels and serum I-FABP levels in patients at T1 compared to T0 (n = 23 samples/group). N Correlation analysis between preoperative fecal MA levels and serum D-lactate levels in patients at T1 compared to T0 (n = 23 samples/group). Scale bar, 100 μm. The statistical tests employed included: two-tailed student’s t test, Spearman’s correlation coefficients and Mann–Whitney tests. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Each dot represents data from a single sample ([B, C], [E], and [L–N]). Bar graphs represent mean ± SD. Source data are provided as a Source Data file.