Technology and method to study microbial growth and adhesion to host-related surfaces and the host-microbiota interactions

Abstract

The present invention relates to in vitro adhesion modules that allow growth, stabilization and study of microbial communities that adhere to and colonize host-related surfaces, that mimic transport of chemical compounds across epithelial surfaces and simulate host-microorganism interactions and adaptation. It includes the provision of micromolar amounts of oxygen via the basal side of a mucus layer towards the adhered microorganisms thus establishing the microaerophilic conditions prevailing at the base of a biofilm. It can also include cells, simulating the host, in a chamber on the basal side of a functional layer comprising said mucus layer. The adhesion module of the present invention can be placed between the different compartments of the SHIME—the Simulator of the Human Intestinal Microbial Ecosystem. An extension of the SHIME is made where the duodenum, jejunum and ileum are separately mimicked.

1 . An adhesion module comprising a basal compartment and a luminal compartment separated by a semi-permeable membrane; said membrane having an artificial mucus layer applied on its luminal side; and said module being characterized by having anaerobic conditions at its luminal side and aerobic conditions at its basal side; wherein the basal compartment of the module comprises viable cells and wherein one or more microorganisms are attached to said artificial mucus layer. 
     
     
         2 . The module of  claim 1  wherein the mucus layer comprises at least one mucin and the thickness of the layer is between 1-1000 μm. 
     
     
         3 . The module of  claim 1  wherein the aerobic conditions in the basal compartment of the module results in a partial oxygen pressure at the basal side of the mucus layer of between about 1 and 150 mmHg. 
     
     
         4 . The module of  claim 1 , wherein the semi-permeable membrane allows the transport of components between 1-100000 Da. 
     
     
         5 . The module of  claim 1 , wherein the material of the semi-permeable membrane is selected from the group consisting of polycarbonate, poly(diallyldimethylammonium chloride), polyethylene terephtalate, or polyamide (nylon). 
     
     
         6 . The module of  claim 1  further comprising a supportive porous layer on the basal side of said semi-permeable membrane. 
     
     
         7 . The module of  claim 6  wherein the material of the supportive porous layer is selected from the group consisting of poly(diallyldimethylammonium chloride), poly(acrylic acid) or combinations thereof. 
     
     
         8 . (canceled) 
     
     
         9 . The module of  claim 1  wherein the cells comprise Caco-2 cells and/or HT29 cells. 
     
     
         10 . The module of  claim 1  wherein the cells grow in a monolayer with their apical side towards the semi-permeable membrane. 
     
     
         11 . The module as claimed in  claim 1  wherein the apical side of the cells is covered with a protective mesh. 
     
     
         12 . The module as claimed in  claims 1  wherein the module comprises beads, having from inside out, an inner support layer, a semi-permeable membrane and an artificial mucus layer. 
     
     
         13 . The module as claimed in  claim 12 , wherein the beads have a radius between 500 μm and 1 cm. 
     
     
         14 . The module of  claim 12  wherein the inner support layer comprises biochemical slow release oxygen components. 
     
     
         15 . The module of  claim 12  wherein the inner support layer is covered with an intermediate layer containing biochemical slow release oxygen components. 
     
     
         16 . The module as claimed in  claim 1  wherein the surface area of the artificial mucus layer is between 1 cm 2  and 200 cm 2 . 
     
     
         17 . The module as claimed in  claim 1  wherein the surface area of the artificial mucus layer is between about 1 cm 2  and 1000 cm 2 . 
     
     
         18 - 31 . (canceled)