Blood brain barrier model and methods of making and using the same

Abstract

Provided herein is an in vitro model of the blood brain barrier. In some embodiments, the model includes: an endothelial cell layer, and brain tissue layer comprising neuronal cells, and optionally one or more of astrocytes, pericytes, oligodendrocytes, and microglia. In some embodiments, the model further comprises a porous membrane between said endothelial cell layer and the neuronal cell layer. A microfluidic device comprising the same and methods of use thereof are also provided.

That which is claimed is: 
     
         1 . An in vitro model of a blood brain barrier, said model comprising six cell types of:
 astrocytes, pericytes, endothelial cells, neuronal cells, oligodendrocytes and microglia,   wherein said model is in the form of a spheroid comprising said six cell types.   
     
     
         2 . The in vitro model of  claim 1 , wherein the neuronal cells comprise primary neuronal cells or neuronal progenitor cells. 
     
     
         3 . The in vitro model of  claim 1 , wherein the endothelial cells comprise primary endothelial cells or endothelial progenitor cells. 
     
     
         4 . The in vitro model of  claim 1 , wherein the astrocytes comprise primary astrocytes or astrocyte progenitor cells. 
     
     
         5 . The in vitro model of  claim 1 , wherein the pericytes comprise primary pericytes or pericyte progenitor cells. 
     
     
         6 . The in vitro model of  claim 1 , wherein the neuronal cells, endothelial cells, astrocytes and/or pericytes are human. 
     
     
         7 . The in vitro model of  claim 1 , wherein said endothelial cells are microvascular endothelial cells. 
     
     
         8 . The in vitro model of  claim 1 , wherein said endothelial cells are human brain microvascular endothelial cells. 
     
     
         9 . The in vitro model of  claim 1 , wherein the six cell types are present in the spheroid in an amount of 30% endothelial cells, 15% pericytes, 15% astrocytes, 15% oligodendrocytes, 5% microglia, and 20% neurons. 
     
     
         10 . The in vitro model of  claim 1 , wherein said spheroid expresses tight junctions and/or adherens junctions. 
     
     
         11 . A method of screening a compound for passage through a blood brain barrier, comprising:
 providing the in vitro model of  claim 1 ,   applying the compound to the model, and   detecting penetration of the compound through the spheroid,   
       to thereby detect passage of the compound through the blood brain barrier. 
     
     
         12 . A method for determining a physiological response to a compound by a blood brain barrier, comprising:
 providing the in vitro model of  claim 1 ,   applying the compound to the model, and   detecting a physiological response from the spheroid,   
       to thereby determine the physiological response to the compound by the blood brain barrier. 
     
     
         13 . The method of  claim 12 , wherein the physiological response comprises damage, scar tissue formation, infection, cell proliferation, cell migration, burn, cell death, marker release, and/or change in gene expression. 
     
     
         14 . A microfluidic device comprising the in vitro model of  claim 1 , wherein the spheroid is in fluid contact or communication with a liquid. 
     
     
         15 . A method of screening a compound for passage through a blood brain barrier, comprising:
 providing the microfluidic device of  claim 11 ,   applying the compound to the model, and   detecting penetration of the compound through the spheroid,   
       to thereby detect passage of the compound through the blood brain barrier. 
     
     
         16 . A method for determining a physiological response to a compound by a blood brain barrier, comprising:
 providing the microfluidic device of  claim 11 ,   applying the compound to the model, and   detecting a physiological response from the spheroid,   
       to thereby determine the physiological response to the compound by the blood brain barrier. 
     
     
         17 . The method of  claim 16 , wherein the physiological response comprises damage, scar tissue formation, infection, cell proliferation, cell migration, burn, cell death, marker release, and/or change in gene expression.