	US 12,188,051 B2
	Compositions and methods for obtaining vascularized human intestinal organoid tissue, and related uses thereof
Emily Holloway, Ann Arbor, MI (US); and Jason Spence, Ann Arbor, MI (US)
Assigned to The Regents of the University of Michigan, Ann Arbor, MI (US)
Filed by The Regents of the University of Michigan, Ann Arbor, MI (US)
Filed on May 27, 2021, as Appl. No. 17/332,532.
Claims priority of provisional application 63/030,621, filed on May 27, 2020.
Prior Publication US 2021/0371815 A1, Dec. 2, 2021
Int. Cl. C12N 5/071 (2010.01)

CPC C12N 5/069 (2013.01) [C12N 2501/11 (2013.01); C12N 2501/115 (2013.01); C12N 2501/155 (2013.01); C12N 2501/16 (2013.01); C12N 2501/165 (2013.01); C12N 2501/999 (2013.01); C12N 2513/00 (2013.01)]

9 Claims

1. A method, comprising:

culturing hindgut spheroid tissue in vitro, wherein the culturing results in differentiation of the hindgut spheroid tissue into tissue comprising vascularized human intestinal organoid tissue,

wherein the culturing of the hindgut spheroid tissue in vitro comprises the following four successive culturing steps:

a) incubating the hindgut spheroid tissue with a media comprising EGF, Noggin and R-Spondin2 (ENR media), wherein EGF is epidermal growth factor;

b) incubating the result of step a) with ENR media and vascular endothelial growth factor (VEGF);

c) incubating the result of step b) with EGF, bone morphogenic protein 4 (BMP4) and basic fibroblast growth factor (bFGF); and

d) incubating the result of step c with EGF and VEGF.