Blood-Brain Barrier Model

Abstract

A method of creating a multicellular blood-brain barrier model is disclosed. In one embodiment, the method comprises culturing primary brain microvascular endothelial cells or embryonic stem cell-derived endothelial cells upon a permeable support in the presence of neural progenitor cells.

1 . A method of creating a multicellular blood-brain barrier model, comprising the step of: 
 culturing brain microvascular endothelial cells upon a permeable support in the presence of neural progenitor cells, wherein the cultured neural progenitor cells differentiate into mixtures of astrocytes, neurons, and oligodendrocytes such that a multicellular blood-brain barrier model is created.    
   
   
       2 . The method of  claim 1  wherein the endothelial cells are isolated from mammalian brain capillaries.  
   
   
       3 . The method of  claim 1  wherein the endothelial cells are derived from isolated embryonic stem cells.  
   
   
       4 . The method of  claim 1  wherein the endothelial cells form a monolayer wherein the cells are confluent and express an initial TEER of 20-50 Ohmxcm 2  before exposure to the neural cells.  
   
   
       5 . The method of  claim 4  wherein the TEER greater than 100 Ohmxcm 2  after exposure to the neural cells.  
   
   
       6 . The method of  claim 5  wherein the TEER is greater than 200 Ohmxcm 2  after exposure to the neural cells.  
   
   
       7 . The method of  claim 1  wherein the neural progenitor cells are isolated from mammalian cortices.  
   
   
       8 . The method of  claim 7  wherein the neural cells are digested with Accutase ™.  
   
   
       9 . The method of  claim 1  wherein the neural progenitor cells are grown as free-floating neurospheres before exposure to the endothelial cells.  
   
   
       10 . The method of  claim 1  wherein the neural progenitor cells are pre-differentiated before exposure to endothelial cells.  
   
   
       11 . The method of  claim 1  wherein the neural cells are removed after the endothelial cells are confluent and express a TEER of at least 100 Ohmxcm 2 .  
   
   
       12 . A blood-brain barrier model created by the method of  claim 1 .  
   
   
       13 . A blood-brain barrier model comprising three components within a liquid-containing vessel, 
 wherein the first component comprises a confluent layer of brain microvacsular endothelial cells or embryonic stem cell-derived endothelial cells,    the second component comprises a permeable membrane support, wherein the first component forms a layer on the second component,    and the third component comprises either (a) undifferentiated neural progenitor cells that are differentiated after contact with the first component to be a mixture of astrocytes, neurons and oligodendrocytes or (b) neural progenitor cells that have been predifferentiated before contact with the first component to be a mixture of astrocytes, neurons and oligodendrocytes,    wherein the first and second components form a barrier between a top and a bottom chamber of the vessel and the third component is placed in the bottom chamber of the vessel.    
   
   
       14 . The model of  claim 13  wherein the endothelial cells are isolated from mammalian brain capillaries.  
   
   
       15 . The model of  claim 13  wherein the endothelial cells form a monolayer wherein the cells are confluent and express an initial TEER of 20-50 Ohmxcm 2  before exposure to the neural cells.  
   
   
       16 . The model of  claim 13  wherein the TEER is greater than 100 Ohmxcm 2  after exposure to the neural cells.  
   
   
       17 . The model of  claim 13  wherein the TEER is greater than 200 Ohmxcm 2  after exposure to the neural cells.  
   
   
       18 . The model of  claim 13  wherein the neural progenitor cells are isolated from mammalian cortices.  
   
   
       19 . The model of  claim 18  wherein the cells are digested with Accutase™.  
   
   
       20 . The model of  claim 13  wherein the neural progenitor cells are grown as free-floating neurospheres before exposure to the endothelial cells.  
   
   
       21 . The model of  claim 13  wherein the neural progenitor cells are pre-differentiated before exposure to the endothelial cells.  
   
   
       22 . The model of  claim 13  wherein the third component has been removed.  
   
   
       23 . A method of analyzing the blood-brain barrier permeability characteristics of a model compound, comprising the steps of: 
 a) exposing a model compound to the blood-brain barrier model of  claim 13  and,    b) measuring the permeability of the barrier model to the compound.    
   
   
       24 . The method of  claim 23  wherein the measurement is by determination of compound concentration in the top and bottom chambers of the vessel.