Coordinated differentiation of human intestinal organoids with functional enteric neurons and vasculature
Charlie J. Childs, Holly M. Poling, Kevin Chen, Yu-Hwai Tsai, Angeline Wu, Abigail Vallie, Madeline K. Eiken, Sha Huang, Caden W. Sweet, Ryan Schreiner, Zhiwei Xiao, Ryan C. Spencer, Samantha A. Paris, Ansley S. Conchola, Jonathan Villanueva, Meghan F. Anderman, Emily M. Holloway, Akaljot Singh, Roman J. Giger, Maxime M. Mahé, Claudia Loebel, Michael A. Helmrath, Katherine D. Walton, Shahin Rafii, Jason R. Spence
Abstract
Human intestinal organoids (HIOs) derived from human pluripotent stem cells co-differentiate both epithelial and mesenchymal lineages in vitro but lack important cell types such as neurons, endothelial cells, and smooth muscle, which limits translational potential. Here, we demonstrate that the intestinal stem cell niche factor, EPIREGULIN (EREG), enhances HIO differentiation 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, HIOs mature and demonstrate enteric nervous system function, undergoing peristaltic-like contractions indicative of a functional neuromuscular unit. HIOs also form functional vasculature, demonstrated in vitro using microfluidic devices and in vivo following transplantation, where HIO endothelial cells anastomose with host vasculature. These complex HIOs represent a transformative tool for translational research in the human gut and can be used to interrogate complex diseases as well as for testing therapeutic interventions with high fidelity to human pathophysiology. • EREG enhances human PSC-derived intestinal organoid differentiation • Organoids possess epithelium, mesenchyme, smooth muscle, neurons, and endothelium • After transplant, EREG organoids mature with human-like organization and function • EREG organoids show peristaltic-like function and have functional vasculature Childs et al. leverage the human intestinal stem cell niche factor EPIREGULIN (EREG) to enhance PSC-derived human intestinal organoid (HIO) differentiations. EREG-grown HIOs pattern epithelium, mesenchyme, smooth muscle, neurons, and endothelium in a single differentiation. After transplant, HIOs mature further, and functional studies show peristaltic-like function and functional vasculature.