In vitro cell culture mucus systems
AssigneeThe University of North Carolina at Chapel Hill
InventorsNancy L. Allbritton, Yuli Wang
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Abstract
This presently disclosed subject matter relates to an in vitro cell culture comprising a cell monolayer comprising mucus producing cells and a mucus layer, and methods of making and using the same. The methods including culturing mucus producing cells on a cell support structure under conditions to establish a mucus layer on the luminal side of the cell monolayer, thereby producing a live cell construct comprising a cell monolayer comprising mucus producing cells and a mucus layer. The mucus layer can be substantially impenetrable to micro-objects, and have a thickness of about 1 micron to about 1 cm.
Claims14
A method of producing a live cell construct comprising a constitutively-produced and continuous mucus layer, the method comprising:
- (a) culturing stem cells that are capable of differentiating into mucus producing cells on a first surface of a porous membrane under a liquid-liquid interface to form a confluent cell monolayer, the liquid-liquid interface comprising:
- (i) a first liquid medium that is in contact with a luminal surface of the cell mono layer, and
- (ii) a second liquid medium that is in contact with a basal surface of the cell monolayer;
- (b) removing the first liquid medium;
- (c) adding vasoactive intestinal peptide (VIP) to the second liquid medium;
- and
- (d) culturing the cell monolayer to generate the constitutively-produced and continuous mucus layer across the luminal surface of the cell monolayer, wherein the constitutively-produced and continuous mucus layer is substantially impenetrable to micro-objects in a size range from about 1 micron to about 100 microns.
Dependencies are inferred from the claim text.