	US 12,104,174 B2
	Methods relating to intestinal organ-on-a-chip
Donald E. Ingber, Boston, MA (US); Magdalena Kasendra, Boston, MA (US); Alexandra Sontheimer-Phelps, Cambridge, MA (US); and Alessio Tovaglieri, Somerville, MA (US)
Assigned to PRESIDENT AND FELLOWS OF HARVARD COLLEGE, Cambridge, MA (US)
Appl. No. 16/331,718
Filed by PRESIDENT AND FELLOWS OF HARVARD COLLEGE, Cambridge, MA (US)
PCT Filed Sep. 13, 2017, PCT No. PCT/US2017/051296 § 371(c)(1), (2) Date Mar. 8, 2019, PCT Pub. No. WO2018/052953, PCT Pub. Date Mar. 22, 2018.
Claims priority of provisional application 62/393,711, filed on Sep. 13, 2016.
Prior Publication US 2020/0231938 A1, Jul. 23, 2020
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/071 (2010.01); C12N 5/00 (2006.01); G01N 33/50 (2006.01)

CPC C12N 5/0697 (2013.01) [C12N 5/0062 (2013.01); C12N 5/0068 (2013.01); C12N 5/0679 (2013.01); G01N 33/5088 (2013.01); C12N 2501/11 (2013.01); C12N 2501/345 (2013.01); C12N 2501/415 (2013.01); C12N 2502/1323 (2013.01)]

39 Claims

1. A method of providing an in vitro intestinal model system, the method comprising:

a) providing i) an intestinal enteroid or colonoid comprising primary intestinal epithelial cells, ii) a microfluidic culture device, the device comprising a porous membrane having first and second surfaces, said membrane in fluidic communication with a microchannel, said microchannel in fluidic communication with a source of fluid;

b) disrupting said intestinal enteroid or colonoid comprising primary intestinal epithelial cells into enteroid or colonoid fragments;

c) seeding said second surface of said porous membrane with said enteroid or colonoid fragments so as to create seeded primary intestinal epithelial cells;

d) expanding said seeded primary intestinal epithelial cells so as to create a monolayer of the intestinal epithelial cells; and

e) exposing said monolayer to a fluid from said source of fluid at a flow rate, wherein said flow results in differentiation of the monolayer of the intestinal epithelial cells and the formation of intestinal villi comprising differentiated intestinal epithelial cells.