US 12,247,227 B2
Methods relating to intestinal organ-on-a-chip
Donald E. Ingber, Cambridge, MA (US); Magdalena Kasendra, Cambridge, MA (US); Alexandra Sontheimer-Phelps, Cambridge, MA (US); and Alessio Tovaglieri, Cambridge, MA (US)
Assigned to PRESIDENT AND FELLOWS OF HARVARD COLLEGE, Cambridge, MA (US)
Filed by PRESIDENT AND FELLOWS OF HARVARD COLLEGE, Cambridge, MA (US)
Filed on Sep. 6, 2023, as Appl. No. 18/242,731.
Application 18/242,731 is a continuation of application No. 17/539,452, filed on Dec. 1, 2021, granted, now 11,976,304.
Application 17/539,452 is a continuation of application No. 16/331,718, granted, now 12,104,174, previously published as PCT/US2017/051296, filed on Sep. 13, 2017.
Claims priority of provisional application 62/393,711, filed on Sep. 13, 2016.
Prior Publication US 2023/0416691 A1, Dec. 28, 2023
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)] 4 Claims
OG exemplary drawing
 
1. A method of providing an in vitro intestinal model system, the method comprising:
a) providing
i) intestinal endothelial cells,
ii) an intestinal enteroid or colonoid comprising primary intestinal epithelial cells,
iii) a microfluidic culture device, the device comprising a porous membrane having a first surface and a second surface, 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 first surface of said porous membrane with said enteroid or colonoid fragments so as to create seeded primary intestinal epithelial cells;
d) seeding said second surface of said porous membrane with said intestinal endothelial cells;
e) expanding said seeded primary intestinal epithelial cells in a first media comprising Wnt-3A so as to create a monolayer of the intestinal epithelial cells;
f) replacing said first media comprising Wnt-3A and culturing said intestinal epithelial cells with a second media lacking Wnt-3A; and
g) culturing said intestinal endothelial cells in a third media comprising Wnt-3A.