Engineered human pluripotent-stem-cell-derived intestinal tissues with a functional enteric nervous system

Abstract

The enteric nervous system (ENS) of the gastrointestinal tract controls many diverse functions, including motility and epithelial permeability. Perturbations in ENS development or function are common, yet there is no human model for studying ENS-intestinal biology and disease. We used a tissue-engineering approach with embryonic and induced pluripotent stem cells (PSCs) to generate human intestinal tissue containing a functional ENS. We recapitulated normal intestinal ENS development by combining human-PSC-derived neural crest cells (NCCs) and developing human intestinal organoids (HIOs). NCCs recombined with HIOs in vitro migrated into the mesenchyme, differentiated into neurons and glial cells and showed neuronal activity, as measured by rhythmic waves of calcium transients. ENS-containing HIOs grown in vivo formed neuroglial structures similar to a myenteric and submucosal plexus, had functional interstitial cells of Cajal and had an electromechanical coupling that regulated waves of propagating contraction. Finally, we used this system to investigate the cellular and molecular basis for Hirschsprung's disease caused by a mutation in the gene PHOX2B. This is, to the best of our knowledge, the first demonstration of human-PSC-derived intestinal tissue with a functional ENS and how this system can be used to study motility disorders of the human gastrointestinal tract.

Pluripotent stem cell-derived organoids: using principles of developmental biology to grow human tissues in a dish

“…Development of these enteric neurons occurs in parallel with that of the epithelium and mesenchymal cell types. Although PSC-derived human intestinal organoids do not contain enteric nerves, a recent study engineered neurons into organoids by experimentally combining PSC-derived neural crest cells with mid/ hindgut spheroids, resulting in self-organizing human intestinal organoids with a functional enteric nervous system (Workman et al, 2016). This system was used to study Hirschsprung's disease, a gut motility disorder resulting from defective enteric neuron development.…”

Ethical issues in human organoid and gastruloid research

“…Nevertheless, a number of studies have begun to explore a higher level of organoid complexity and functionality (Assawachananont et al, 2014;Takebe et al, 2013;Workman et al, 2017). Retinal organoids generated from mouse PSCs can functionally engraft, exhibiting synaptic connections with the retina in host mice (Assawachananont et al, 2014).…”

“…Subsequently, complex vascularized organ buds for kidney, pancreas, intestine, heart, lung and brain have been developed through co-culture of mouse PSC-derived progenitors, endothelial cells and mesenchymal stem cells following engraftment in host mice (Takebe et al, 2015). Innervation of PSC-derived organoids has also recently been achieved (Workman et al, 2017). Specifically, developing PSC-derived intestinal organoids were co-cultured with PSC-derived neural crest cells in order to recreate a functional enteric nervous system.…”

Regenerative Medicine: Advances from Developmental to Degenerative Diseases

“…HPSC-derived enteric neurons can also innervate hPSC-derived gut organoids, which represent the three-dimensional models of the gut tissue comprising various cell types present in the gut epithelia as well as the smooth muscle that surrounds it 57 . Combining the enteric neural progenitors with gut organoids led to the formation of enteric ganglia and innervation of the smooth muscle, thus allowing the control of peristalsis 57 .…”

“…Combining the enteric neural progenitors with gut organoids led to the formation of enteric ganglia and innervation of the smooth muscle, thus allowing the control of peristalsis 57 .…”