In vitro gastrointestinal model comprising lamina propria-derived cells

Abstract

An in vitro microfluidic gut-on-chip is described herein that mimics the structure and at least one function of specific areas of the gastrointestinal system in vivo. In particular, a multicellular, layered, microfluidic culture is described, allowing for interactions between lamina propria-derived cells and gastrointestinal epithelial cells and endothelial cells. This in vitro microfluidic system can be used for modeling inflammatory gastrointestinal tissue, e.g., Crohn's disease, colitis and other inflammatory gastrointestinal disorders. These multicellular, layered microfluidic gut-on-chip further allow for comparisons between types of gastrointestinal tissues, e.g., small intestinal deuodejeum, small intestinal ileium, large intestinal colon, etc., and between disease states of gastrointestinal tissue, i.e. healthy, pre-disease and diseased areas. Additionally, these microfluidic gut-on-chips allow identification of cells and cellular derived factors driving disease states and drug testing for reducing inflammation.

1 . A microfluidic device comprising a microfluidic channel and a membrane in contact with said microfluidic channel, said membrane comprising an extracellular matrix (ECM) comprising one or more extracellular proteins and affinity reagents, said affinity reagents bound to said ECM and configured to stimulate T helper cells. 
     
     
         2 - 6 . (canceled) 
     
     
         7 . The microfluidic device of  claim 1 , wherein said affinity reagents are covalently attached within said microfluidic device. 
     
     
         8 - 14 . (canceled) 
     
     
         15 . The microfluidic device of  claim 1 , wherein said one or more extracellular matrix proteins comprise an overlay, said overlay trapping said affinity reagents. 
     
     
         16 . (canceled) 
     
     
         17 . The microfluidic device of  claim 1 , wherein said affinity reagents comprises antibodies or binding fragments thereof. 
     
     
         18 . The microfluidic device of  claim 17 , wherein said antibodies are anti-CD3 antibodies. 
     
     
         19 . The microfluidic device of  claim 17 , wherein said antibodies are anti-CD28 antibodies. 
     
     
         20 . The microfluidic device of  claim 17 , wherein said antibodies are a combination of anti-CD3 antibodies and anti-CD28 antibodies. 
     
     
         21 - 23 . (canceled) 
     
     
         24 . The microfluidic device of  claim 1 , further comprising T helper cells. 
     
     
         25 - 26 . (canceled) 
     
     
         27 . The microfluidic device of  claim 24 , wherein said T helper cells comprise TH1 cells. 
     
     
         28 . The microfluidic device of  claim 24 , wherein said T helper cells comprise TH9 cells. 
     
     
         29 . The microfluidic device of  claim 1 , wherein said device further comprises soluble antigen. 
     
     
         30 - 73 . (canceled) 
     
     
         74 . A microfluidic device, wherein said microfluidic device comprises one or more microfluidic channels, a membrane, said membrane comprising a top surface and a bottom surface, and a co-culture of gastrointestinal epithelial cells and lamina propria-derived cells located on said top surface of said membrane, and endothelial cells located on said bottom surface of said membrane. 
     
     
         75 . The microfluidic device of  claim 74 , wherein said membrane is semi-permeable. 
     
     
         76 . The microfluidic device of  claim 74 , wherein said lamina propria-derived cells comprise stromal cells and immune cells. 
     
     
         77 . The microfluidic device of  claim 76 , wherein said immune cells are selected from the group consisting of lymphocytes, mononuclear cells, macrophages, immature dendritic cells, mature dendritic cells, eosinophils, basophils, mast cells and combinations thereof. 
     
     
         78 . The microfluidic device of  claim 74 , wherein said top surface of said membrane comprises at least one extracellular matrix protein. 
     
     
         79 . The microfluidic device of  claim 74 , wherein said lamina propria-derived cells were obtained from human gastrointestinal tissue selected from the group consisting of healthy tissue, pre-diseased tissue and diseased tissue. 
     
     
         80 . The microfluidic device of  claim 74 , wherein said lamina propria-derived cells were obtained from inflamed human gastrointestinal tissue. 
     
     
         81 . The microfluidic device of  claim 74 , wherein said lamina propria-derived cells were obtained from a human with inflammatory colitis. 
     
     
         82 . The microfluidic device of  claim 74 , further comprising media, wherein said media is located within said microfluidic channels and in contact with said co-culture of said cells. 
     
     
         83 . A microfluidic system comprising a microfluidic device and a co-culture of gastrointestinal epithelial cells, and lamina propria-derived cells, wherein said co-culture is perfused with fluid under flow conditions. 
     
     
         84 . The microfluidic system of  claim 83 , wherein said device further comprises endothelial cells. 
     
     
         85 . The microfluidic system of  claim 83 , wherein said device comprises a membrane, said membrane comprising a top surface and a bottom surface, and one or more microfluidic channels in fluidic communication with a source of said fluid. 
     
     
         86 . The microfluidic system of  claim 85 , wherein said microfluidic device comprises first and second microfluidic channels separated by said membrane. 
     
     
         87 . The microfluidic system of  claim 85 , wherein said top surface of said membrane comprises at least one extracellular matrix protein. 
     
     
         88 . The microfluidic system of  claim 85 , wherein said gastrointestinal cells are located on said top surface of said membrane. 
     
     
         89 . The microfluidic system of  claim 85 , further comprising endothelial cells are on said bottom surface of said membrane. 
     
     
         90 . The microfluidic system of  claim 83 , wherein said lamina propria-derived cells comprise stromal cells and immune cells. 
     
     
         91 . The microfluidic system of  claim 83 , wherein said lamina propria-derived cells comprise resident immune cells of human gastrointestinal tissue. 
     
     
         92 . The microfluidic system of  claim 83 , wherein said lamina propria-derived cells are obtained from gastrointestinal tissue and are selected from the group consisting of healthy tissue, pre-diseased tissue and diseased tissue.