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US 11,767,515 B2Granted
Colonic organoids and methods of making and using same
AssigneeChildren's Hospital Medical Center
InventorsJames M. Wells, Jorge Orlando Munera
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Abstract
Disclosed herein are methods for the in vitro differentiation of a precursor cell into definitive endoderm, which may further be differentiated into a human colonic organoid (HCO), via modulation of signaling pathways. Further disclosed are HCOs and methods of using HCOs, which may be used, for example, for the HCOs may be used to determine the efficacy and/or toxicity of a potential therapeutic agent for a disease selected from colitis, colon cancer, polyposis syndromes, and/or irritable bowel syndrome.
Claims31
A method of inducing formation of a human colon organoid (HCO), comprising the steps of
- (a) contacting a definitive endoderm (DE) with an FGF signaling pathway activator and a WNT signaling pathway activator for a period of time sufficient for said DE to form a mid-hindgut spheroid;
- (b) contacting the mid-hindgut spheroid of step (a) with a BMP activator and an EGF signaling pathway activator for about 1 day to about 3 days;
- and
- (c) contacting the mid-hindgut spheroid of step (b) with an EGF signaling pathway activator without a BMP activator for a period of time sufficient to form said human colon organoid, wherein said human colon organoid expresses SATB2.
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Dependencies are inferred from the claim text.