Back to results
US 11,884,938 B2Granted
Cell culture system
AssigneePresident and Fellows of Harvard College
InventorsDonald E. Ingber, Hyun Jung Kim
Life of this patent
Estimating the term…
Abstract
The embodiments of the invention described herein relate to systems and methods for culturing and/or maintaining intestinal cells, tissues and/or organoids in vitro. The cells, tissues and/or organoids cultured according to the methods and systems described herein can mimic or reproduce natural intestinal epithelial structures and behavior as well as support co-culture of intestinal microflora.
Claims19
A method for culturing intestinal epithelial cells comprising:
- a) providing a microfluidic device comprising a microchannel connected to a fluid source comprising culture media, and a membrane positioned within the microchannel;
- b) introducing intestinal epithelial cells to at least one surface of said membrane;
- c) supplying culture media from the fluid source to the microchannel at a flow rate of less than 500 μL/hr;
- and
- d) culturing the intestinal epithelial cells at least until villi structures develop within 132 to 170 hours, wherein said culturing comprises:
- i) stretching said membrane from 5% to 15% along at least one dimension of the membrane during the culturing step;
- and
- ii) stretching said membrane in a cyclic manner at a rate in the range of 0.01 Hz to 0.15 Hz during the culturing step.
5 more elements
A method for culturing intestinal epithelial cells comprising:
- a) providing a microfluidic device comprising a microchannel connected to a fluid source comprising culture media, and a membrane positioned within the microchannel;
- b) introducing intestinal epithelial cells to at least one surface of said membrane, wherein said intestinal epithelial cells are selected from the group consisting of Caco2 cells;
- HT-29 cells;
- primary small intestine epithelial cells;
- and primary large intestine epithelial cells;
- c) supplying culture media from the fluid source to the microchannel at a flow rate of less than 500 μL/hr;
- and
- d) culturing the intestinal epithelial cells at 37° C. at least until villi structures develop within 132 to 170 hours, wherein said culturing comprises:
- i) stretching said membrane from 5% to 15% along at least one dimension of the membrane during the culturing step;
- and
- ii) stretching said membrane in a cyclic manner at a rate in the range of 0.01 Hz to 0.15 Hz during the culturing step.
8 more elements
Dependencies are inferred from the claim text.