Figure 1

IDH1 mutation enhances the formation of biliary organoids established from murine liver. (A) The left panel shows a representative image of IBOs from wild-type mice (8-weeks old, male) at day 7. The middle and left panels show hematoxylin and eosin (HE) staining of them. Scale bars, 250 μm (middle and left panels) and 25 μm (right panel). (B) Immunohistochemical staining of Ck19 and Sox9 in IBOs. Scale bars, 20 μm. (C) The amount of 2-HG measured by CE-MS in cell extracts from IBOs stably expressing empty vector (EV), wild-type (Wild), and mutant IDH1 (R132C). (n = 4, *P < 0.05, NS not significant). (D) The number of IBOs (>100 μm) at 7 d after plating (n = 4, 3,000 cells per group, *P < 0.05) and their representative images. Scale bars, 250 μm. The assay was performed at passage 4. (E) Organoid-forming efficiency of the indicated IBOs during serial passage (P5–7) after puromycin selection. The number of organoids (>100 μm) at 7 d after plating (n = 4, 3,000 cells per group, *P < 0.05). (F) IBOs established from wild-type mice treated with 10 mM 2-HG ( + ) or vehicle (−) upon serial passage. The number of organoids (>100 μm) at 7 d after plating (n = 5, 3,000 cells per group, *P < 0.05, NS not significant). (G) 2-HG levels of mut-IBOs treated with 20 μM AGI-5198 (+) or DMSO vehicle (−). (H,I) The number (>100 μm) of mut-IBOs (H) and wt-IBOs (I) treated with 20 μM AGI-5198 (+) or DMSO vehicle at 7 d after plating (n = 4, 10,000 cells per group, *P < 0.05).