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. Author manuscript; available in PMC: 2019 Aug 15.
Published in final edited form as: Brain Res. 2018 Aug 15;1693(Pt B):165–168. doi: 10.1016/j.brainres.2018.04.015

FIGURE 1. Generation of Human Intestinal Organoids with an Enteric Nervous System.

FIGURE 1

Schematic for generating human intestinal organoids (HIO) and vagal neural crest cells (NCC) through directed differentiation of human pluripotent stem cells. Activin A mediates efficient differentiation into definitive endoderm and WNT activation, using CHIR99021, in combination with FGF4 induces midgut endoderm monolayers and free-floating spheroids. FGF and EGF mediate differentiation into NCCs. Retinoic acid posteriorizes the NCCs into vagal NCCs. HIOs and NCCs are generated separately, combined by low-speed centrifugation, embedded in Matrigel and grown for 4 weeks in vitro.