Figure 2: Barrett’s stem cell pedigree recapitulates intestinal metaplasia.
From: Mutational spectrum of Barrett’s stem cells suggests paths to initiation of a precancerous lesion
(a) Derivation of stem cell pedigree from pool of colonies (left) by selecting individual colony for replating and expansion (right). Scale bar, 200 μm. n=12 biological replicates. (b) Clonogenicity assay for BESC at passage 5 and passage 12 determined by plating 2,000 cells and counting colony formation following Rhodamine red staining. Histogram depicting colony counts. Error bars, s.d. n=3 biological replicates. (c) Left, schematic diagram of induced differentiation of stem cell pedigree in ALI culture. Right, histological section of differentiated p5 and p12 Barrett’s stem cells with Alcian blue-positive goblet cells. Scale bar, 100 um. n=12 biological replicates. (d) Upper, Alcian blue staining of ALI culture differentiation of EsoSC, BESC and GSC, bottom, immunostainings of section of BESC-derived epithelium following ALI differentiation showed distribution of multiple types of cells positive for CHGA (endocrine cells), HD6 (Paneth cells) and TFF3 (goblet cells). Scale bar, 50 μm. n=12 biological replicates. (e) PCA of whole-genome expression data sets from EsoSC, BESC and GSC before and after ALI differentiation as indicated. (f) Expression heatmap of differentially expressed genes in ALI cultures as indicated.
