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. 2017 Nov 15;68(1):49–61. doi: 10.1136/gutjnl-2017-314817

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Altered DNA methylation in organoids derived from gastric heterotopia. (A) Endoscopic image of rectal mucosa. Numbers indicate gastric heterotopic region (=1), mucosal ulcer (=2) and healthy, unaffected rectal mucosa (=3). (B) (I) Sections of healthy gastric tissue (STO), gastric heterotopic tissue in the rectum (GHR) and adjacent healthy rectum (REC). Shown are H&E staining as well as immunohistochemistry staining for hindgut marker CDX2 and gastric marker KRT7. (II) Intestinal epithelial organoids (IEOs) derived from stomach, gastric heterotopic and healthy rectal tissue. (C) Multidimensional scaling plot of genome-wide DNA methylation of different IEOs from the same donor. Samples were passages 3 and 5, indicated by superscript number next to the sample. STO-organoids were grown in gastric medium, REC-organoids in intestinal medium and GHR-organoids in both gastric (P3 and P5) and intestinal medium (P5). (D) Methylation levels of CpGs located in the genes of CDX2 (left) and KRT7 (right). Box plot of n=2–3 IEO per group. (E) Pyrosequencing data showing percentage of methylation in the genes of SATB2-AS1 and GATA2 in heterotopic and rectal IEOs. Data shown as mean+SD of n=2 per group. See also online supplementary figure S8. G, gastric heterotopic organoids; R, rectal organoids; S, stomach-derived organoids.