US 11,053,477 B2
Methods and systems for converting precursor cells into gastric tissues through directed differentiation
James Macormack Wells, Cincinnati, OH (US); and Kyle William McCracken, Cincinnati, OH (US)
Assigned to Children's Hospital Medical Center, Cincinnati, OH (US)
Filed by Children's Hospital Medical Center, Cincinnati, OH (US)
Filed on Nov. 13, 2018, as Appl. No. 16/188,597.
Application 16/188,597 is a continuation of application No. 15/312,939, granted, now 10,174,289, previously published as PCT/US2015/032626, filed on May 27, 2015.
Claims priority of provisional application 62/003,719, filed on May 28, 2014.
Prior Publication US 2019/0153397 A1, May 23, 2019
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/071 (2010.01)
CPC C12N 5/0697 (2013.01) [C12N 5/0679 (2013.01); C12N 2501/11 (2013.01); C12N 2501/113 (2013.01); C12N 2501/115 (2013.01); C12N 2501/119 (2013.01); C12N 2501/155 (2013.01); C12N 2501/16 (2013.01); C12N 2501/385 (2013.01); C12N 2501/415 (2013.01); C12N 2501/727 (2013.01); C12N 2506/02 (2013.01); C12N 2506/45 (2013.01)] 18 Claims
OG exemplary drawing
 
1. 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 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.