Development of Colonic Organoids Containing Enteric Nerves or Blood Vessels from Human Embryonic Stem Cells

Abstract

The increased interest in organoid research in recent years has contributed to an improved understanding of diseases that are currently untreatable. Various organoids, including kidney, brain, retina, liver, and spinal cord, have been successfully developed and serve as potential sources for regenerative medicine studies. However, the application of organoids has been limited by their lack of tissue components such as nerve and blood vessels that are essential to organ physiology. In this study, we used three-dimensional co-culture methods to develop colonic organoids that contained enteric nerves and blood vessels. The development of enteric nerves and blood vessels was confirmed phenotypically and genetically by the use of immunofluorescent staining and Western blotting. Colonic organoids that contain essential tissue components could serve as a useful model for the study of colon diseases and help to overcome current bottlenecks in colon disease research.

Organoids in gastrointestinal diseases: from experimental models to clinical translation

“…62 Current models have also utilised the PSC system that can be differentiated towards intestinal organoids, as well as cells of the ENS such as neural crest cells (NCCs). [63][64][65] NCCs that were cocultured with developing intestinal organoids migrated into the mesenchyme, differentiated into neurons and glial cells and showed neuronal activity. 63 ENS-containing intestinal organoids formed neuroglial structures similar to a myenteric and submucosal plexus in vivo.…”

“… 73 Indeed, several attempts have been made to introduce iPSC-derived endothelial cells into GI organoids. 65 74 75 Interestingly, a recent study uncovered a population of endothelial cells which are present early in iPSC differentiation towards intestinal organoids that declines over time in culture. This study further developed a method to expand and maintain this endothelial cell popoulation within intestinal organoids as an alternative approach for classical vascularised organoid models.…”

Technical advances in pluripotent stem cell-derived and tumorigenic organoids

“…The development of colonic organoids from PSCs has recently advanced. Park et al [35] reported a method that generated colonic organoids containing enteric nerves or blood vessels by co-culturing nerve cells or endothelial cells with colonic organoids. The NCCs and endothelial cells used in that study were all differentiated from the same hPSC line as the colonic organoids, and integration into organoids was performed after the hindgut was defined from the definitive endoderm.…”

Shaping axial identity during human pluripotent stem cell differentiation to neural crest cells

“…When combined, vagal NC progenitors generated glia and neurons of the ENS which integrated into the smooth muscle layer of the HIO and were found to promote cell proliferation in intestinal crypts and gene expression indicative of intestinal cell development. More recent work has described the generation of assembloid cultures containing colonic intestinal tissue with ENS and blood vessels; a model perhaps more suited to diseases which impact the distal part of the GI tract, including Crohn's and Hirschsprung's disease [ 88 ].…”