Figure 1.

Schematic of differentiation into human intestinal and colonic organoids. Human pluripotent stem cells are differentiated into definitive endoderm (DE) with co-developing mesoderm by treatment with Activin A for 3 days. The resulting DE is then treated with WNT and FGF4 for patterning into mid/hindgut endoderm with posterior mesoderm. After 4 days of treatment with WNT/FGF4, floating spheroids will be evident. These floating spheroids can be plated in a Matrigel bubble and then BMP signaling can either be inhibited for 3 days to generate HIOs or activated for 3 days to generate HCOs with increasing numbers of goblet cells present in HCOs. Human pluripotent stem cells are depicted in green. Definitive endoderm and DE derived cells are depicted in yellow. Mesoderm and mesoderm derived cells are depicted in red. Goblet cells within HIOs and HCOs are depicted in cyan.