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US 10,174,289 B2Granted
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.
Claims14
A method of inducing formation of a gastric tissue, comprising the steps of:
- a) contacting a culture containing a mammalian definitive endoderm cell 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 spheroids express SOX2, PDX1, HNF1β, 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 cell types selected from one or more of antral mucous cells and endocrine cells.
Dependencies are inferred from the claim text.