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US 11,066,650 B2Granted
Methods for the in vitro manufacture of gastric fundus tissue and compositions related to same
AssigneeChildren's Hospital Medical Center
InventorsJames Wells, Kyle McCracken
Life of this patent
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Abstract
The instant disclosure relates to methods for converting mammalian definitive endoderm (DE) cells into specific tissue(s) or organ(s) through directed differentiation. In particular, the disclosure relates to formation of gastric fundus tissue and/or organoids formed from differentiated definitive endoderm.
Claims16
An in vitro method of inducing formation of a gastric fundus tissue comprising a functional fundic cell type, comprising the steps of:
- a) contacting a mammalian definitive endoderm (DE) cell with a wnt activator, an FGF activator, a BMP inhibitor, and retinoic acid, for a first period, wherein said first period is for a length of time sufficient to form a three-dimensional posterior foregut spheroid from said definitive endoderm;
- b) contacting said three-dimensional posterior foregut spheroid with a growth factor, said Wnt activator, an EGF signalling pathway activator, said BMP inhibitor, and retinoic acid for a second period, wherein said second period is for a length of time sufficient to induce a fundic lineage comprising human fundic-type gastric organoids (hFGOs);
- c) culturing said hFGOs of step b) with said wnt activator and said EGF activator for a third period;
- d) culturing said hFGOs of step c) with said wnt activator, said EGF activator, and FGF10 for a fourth period;
- e) contacting said hFGOs of step d) with a MEK inhibitor for a fifth period, wherein said fifth period is for a period of time sufficient to form said gastric fundus tissue comprising a functional fundic cell type.
2 more elements
An in vitro method of forming a functional fundic cell type comprising the steps of:
- a) contacting an hFGO with a wnt activator and an EGF activator for a first period, and
- b) contacting the hFGO of step a) with a MEK inhibitor for a second period, wherein said first and second periods are carried out for a period of time sufficient to form a functional fundic cell type;
- wherein said hFGO is obtained by contacting a three-dimensional posterior foregut spheroid with a growth factor, a wnt activator, an EGF activator, a BMP inhibitor, and retinoic acid for a period of time sufficient to convert said three-dimensional posterior foregut spheroid to said hFGO;
- wherein said three-dimensional posterior foregut spheroids are obtained by contacting a mammalian DE cells with a wnt activator, an FGF activator, a BMP inhibitor, and retinoic acid.
1 more element
Dependencies are inferred from the claim text.