Methods and systems for converting precursor cells into gastric tissues through directed differentiation
AssigneeChildren's Hospital Medical Center
InventorsJames Macormack Wells, Kyle William McCracken
Life of this patent
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Abstract
Disclosed are methods of inducing formation of a gastric cells and/or a gastric tissue, such as in the form of a gastric organoid. The formation of gastric cells and/or tissue may be carried out by the activating and/or inhibiting of one or more signaling pathways within a precursor cell. Also disclosed are methods for using the disclosed gastric cells, gastric tissues, and/or gastric organoids derived from precursor cells.
Claims18
A method of inducing formation of a gastric tissue, comprising the steps of:
- a) contacting a culture containing a mammalian definitive endoderm with an FGF signaling pathway activator, a Wnt signaling pathway activator, and a BMP inhibitor for at least two days or for a period of time sufficient to form SOX2+ foregut cells;
- b) contacting said SOX2+ foregut cells of step (a) with retinoic acid for a period of time sufficient to form three-dimensional foregut spheroids, wherein said three-dimensional foregut spheroids express SOX2, PDX1, HNF1beta, and HNF6;
- c) contacting said three-dimensional foregut spheroids of step (b) with retinoic acid, EGF and a BMP inhibitor for a period of time sufficient for formation of a gastric organoid, wherein said gastric organoid is characterized by expression of MUC5AC, gastrin, ghrelin, 5-HT, and ChrA and the presence of a cell type selected from an antral mucous cell, an endocrine cell, and a combination thereof.
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A method of generating a gastric organoid comprising fundus tissue comprising the steps of
- a) generating a definitive endoderm (DE) from a pluripotent stem cell;
- b) contacting said definitive endoderm with an agent that activates the FGF pathway, an agent that activates the WNT pathway, an agent that inhibits the BMP pathway, and retinoic acid to generate a posterior foregut;
- and
- c) contacting said posterior foregut with EGF, retinoic acid, an agent that activates the WNT pathway, and an agent that inhibits the BMP pathway to generate a fundus tissue.
A method of generating a gastric organoid comprising antrum tissue comprising the steps of
- a) generating a definitive endoderm (DE) from a pluripotent stem cell;
- b) contacting said definitive endoderm with an agent that activates the FGF pathway, an agent that activates the WNT pathway, an agent that inhibits the BMP pathway, and retinoic acid to generate a posterior foregut;
- and
- c) contacting said posterior foregut with EGF, retinoic acid, and an agent that inhibits the BMP pathway to generate an antrum tissue.
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Dependencies are inferred from the claim text.