Coordinated differentiation of human intestinal organoids with functional enteric neurons and vasculature

Abstract

AbstractHuman intestinal organoids (HIOs) derived from human pluripotent stem cells co-differentiate both epithelial and mesenchymal lineagesin vitrobut lack important cell types such as neurons, endothelial cells, and smooth muscle. Here, we report anin vitromethod to derive HIOs with epithelium, mesenchyme, enteric neuroglial populations, endothelial cells, and organized smooth muscle in a single differentiation, without the need for co-culture. When transplanted into a murine host, these populations expand and organize to support organoid maturation and function. Functional experiments demonstrate enteric nervous system function, with HIOs undergoing peristaltic-like contractions, suggesting the development of a functional neuromuscular unit. HIOs also form functional vasculature, demonstratedin vitrousing microfluidic devices to introduce vascular-like flow, andin vivofollowing transplantation, where HIO endothelial cells anastomose with host vasculature. Collectively, we report anin vitromodel of the human gut that simultaneously co-differentiates epithelial, stromal, endothelial, neural, and organized muscle populations.