Deriving functional human enteroendocrine cells from pluripotent stem cells

Abstract

Enteroendocrine cells (EECs) are a minor cell population in the intestine yet they play a major role in digestion, satiety, and nutrient homeostasis. Recent development of human intestinal organoid models include EECs, but their rarity makes it difficult to study their formation and function. Here we used the EEC-inducting property of the transcription factor NEUROG3 in human pluripotent stem cell (PSC)-derived human intestinal organoids (HIOs) and colonic organoids (HCOs) to promote EEC development in vitro. An 8 hour pulse of NEUROG3 expression induced expression of known target transcription factors and after 7 days organoids contained up to 25% EECs in the epithelium. EECs expressed a broad array of human hormones at the mRNA and /or protein level, including MOTILIN, SOMATOSTATIN, NEUROTENSIN, SECRETIN, SUBSTANCE P, SEROTONIN, VIP, OXYNTOMODULIN, GLP-1 and INSL5. EECs secreted several hormones including GIP, GHRELIN, GLP-1, and OXYNTOMODULIN. Injection of glucose into the lumen of organoids caused an increase in both GIP secretion and K-cell number. Lastly, we observed formation of all known small intestinal EEC subtypes following transplantation and growth of HIOs in mice.