Extended Data Figure 6. Induction of genes during development of hGOs that mark specific differentiated antral cell types.

a, qPCR analyses of cell lineage differentiation marker expression at several stages throughout the gastric organoid differentiation protocol (d0, d3, d6, d9, d20, d27, and d34) and d34 human intestinal organoids (hIO). Beginning at day 27, hGOs robustly induced genes expressed in differentiated cell types including surface mucous cells (MUC5AC, TFF1, TFF3, and GKN1) and antral gland cells (TFF2). HGOs do not up-regulate expression of markers found in fundic lineages such as parietal cells (ATP4A and ATP4B) and chief cells (MIST1) or intestinal goblet cells (MUC2). Expression levels are normalized to d3 DE cultures. n=3 biological replicates per time point. b, Muc5AC-expressing surface mucous cells in the late fetal (E18.5) mouse antrum are not yet confined to a pit region and are more broadly distributed through the antral epithelium. Further, these pit cells exhibit high amounts of intracellular mucin staining, similar to 34-day hGOs. c, Global gene expression profiling of 34-day hGOs was performed using RNA-seq, and data were compared to published RNA-seq datasets from human tissues. Hierarchical clustering revealed that hGOs closely resemble human fetal stomach tissue but not human fetal intestine. Error bars represent standard deviation.