Generation of stomach tissue from mouse embryonic stem cells

Abstract

Successful pluripotent stem cell differentiation methods have been developed for several endoderm-derived cells, including hepatocytes, β-cells and intestinal cells. However, stomach lineage commitment from pluripotent stem cells has remained a challenge, and only antrum specification has been demonstrated. We established a method for stomach differentiation from embryonic stem cells by inducing mesenchymal Barx1, an essential gene for in vivo stomach specification from gut endoderm. Barx1-inducing culture conditions generated stomach primordium-like spheroids, which differentiated into mature stomach tissue cells in both the corpus and antrum by three-dimensional culture. This embryonic stem cell-derived stomach tissue (e-ST) shared a similar gene expression profile with adult stomach, and secreted pepsinogen as well as gastric acid. Furthermore, TGFA overexpression in e-ST caused hypertrophic mucus and gastric anacidity, which mimicked Ménétrier disease in vitro. Thus, in vitro stomach tissue derived from pluripotent stem cells mimics in vivo development and can be used for stomach disease models.

The hope and the hype of organoid research

“…Analysis of lung organoids has taught us that FGF signalling specifies human anterior foregut and SHH specifies lung fate (Dye et al, 2015), consistent with the role of these pathways in mouse lung growth (Morrisey and Hogan, 2010). Similarly, by differentiating PSCs into stomach organoids, we have learned that Wnt/β-catenin signalling is essential to drive the production of both human and mouse stomach acid-producing cells (Noguchi et al, 2015;McCracken et al, 2017). These pioneering studies are already revolutionising the way we study human development and show that organoid cultures have the potential to advance the field of human development in an unprecedented manner.…”

Stomach development, stem cells and disease

“…These aspects of pathobiology have successfully been reproduced in antral organoid cultures derived from human ESCs (McCracken et al, 2014) or primary human corpus specimens (Bartfeld et al, 2015). Mouse organoid cultures have also been used to assess parietal cell function and repair following cell damage induced by a two-photon laser (Schumacher et al, 2015) and to replicate features of Menetriere disease (Noguchi et al, 2015), which is a rare premalignant disease of the stomach. These advances emphasize the value of insights from developmental biology in tissue engineering and in vitro disease modeling.…”

“…Their recapitulation of these pathway activities in mouse ESCs, followed by Barx1 activation in mesenchymal cells, yielded stomach organoids that resemble either the antrum or corpus, with the latter containing mature parietal and chief cells (Noguchi et al, (McCracken et al, 2014). (B) After induction of definitive endoderm in murine ESCs, DKK1, SHH and knockout serum replacement (KSR) are added to small spheroids, followed by 3D culture in medium containing FGF10, WNT3A, Noggin and RSPO1 to promote corpus organoid differentiation (Noguchi et al, 2015). (C) Single human gastric epithelial cells, isolated by fluorescent cell sorting, are exposed to EGF, Noggin, RSPO1, Wnt, FGF10 and gastrin (ENRWFG), followed by removal of Wnt, to induce stomach organoids (Bartfeld et al, 2015).…”

Modeling infectious diseases and host-microbe interactions in gastrointestinal organoids

“…In humans, loss of parietal cells is a very early step in the process of gastric carcinogenic. Parietal cells have been refractory to previous culture attempts, but organoid cultures derived from mESCs (Noguchi et al, 2015), as well as short-term cultures of murine and human ASCs, harbor parietal cells (Schumacher et al, 2015a). Shh is induced in wild-type murine ASCderived organoids infected with H. pylori, whilst organoids derived from mice with a parietal cell-specific deletion of Shh were unable to upregulate Shh, indicating that H. pylori infection induces Shh transcription specifically in parietal cells.…”