Blood-Brain Barrier Model

1 . A method of creating a multicellular blood-brain barrier model, comprising the step of:

(a) 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 Ohm×cm 2 before exposure to the neural cells.

5 . The method of claim 4 wherein the TEER greater than 100 Ohm×cm 2 after exposure to the neural cells.

6 . The method of claim 5 wherein the TEER is greater than 200 Ohm×cm 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 at least one enzyme to dissociate the cells.

9 . The method of claim 1 wherein the neural progenitor cells are grown as free-floating neurospheres before differentiation.

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 Ohm×cm 2 .

12 . A method of creating a multicellular blood-brain barrier model, comprising the step of:

(a) culturing brain microvascular endothelial cells upon a permeable support in the presence of multipotent neural progenitor cells, wherein the endothelial cells form a monolayer wherein the cells are confluent and express an initial transendothelial electrical resistance (TEER) of 20-50 Ohm×cm 2 before exposure to the neural cells, wherein the multipotent neural progenitor cells further differentiate into mixtures of astrocytes, neurons, and oligodendrocytes, wherein the TEER is greater than 100 Ohm×cm 2 after exposure of the endothelial cells to the differentiated neural cells and wherein the model is then capable of a TEER of greater than 100 Ohm×cm 2 for a period of at least 72 hours.

13 . The method of claim 12 wherein the neural progenitor cells are pre-differentiated before exposure to endothelial cells.

14 . A blood-brain barrier model created by the method of claim 1 .

15 . A blood-brain barrier model created by the method of claim 10 .

16 . A blood-brain barrier model created by the method of claim 12 .

17 . A blood-brain barrier model created by the method of claim 13 .

18 . A blood-brain barrier model comprising three components within a liquid-containing vessel,

wherein the first component comprises a confluent layer of brain microvascular 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 pre-differentiated 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.

19 . The model of claim 15 wherein the endothelial cells are isolated from mammalian brain capillaries.

20 . The model of claim 15 wherein the endothelial cells form a monolayer wherein the cells are confluent and express an initial TEER of 20-50 Ohm×cm 2 before exposure to the neural cells.

21 . The model of claim 15 wherein the TEER is greater than 100 Ohm×cm 2 after exposure to the neural cells.

22 . The model of claim 15 wherein the TEER is greater than 200 Ohm×cm 2 after exposure to the neural cells.

23 . The model of claim 15 wherein the neural progenitor cells are isolated from mammalian cortices.

24 . The model of claim 20 wherein the cells are digested with at least one enzyme to dissociate the cells.

25 . The model of claim 15 wherein the neural progenitor cells are grown as free-floating neurospheres before differentiation.

26 . The model of claim 15 wherein the neural progenitor cells are pre-differentiated before exposure to the endothelial cells.

27 . The model of claim 15 wherein the third component has been removed.

28 . A blood-brain barrier model including three components within a liquid-containing vessel, comprising

i) a first component comprising a confluent layer of brain microvascular endothelial cells or embryonic stem cell-derived endothelial cells, wherein the endothelial cells form a monolayer and wherein the cells are confluent and express an initial TEER of 20-50 Ohm×cm 2 before exposure to the neural cells;

ii) a second component comprising a permeable membrane support, wherein the first component forms a layer on the second component, and

iii) a third component comprising 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 pre-differentiated 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, and wherein the TEER is greater than 100 Ohm×cm 2 after exposure of the endothelial cells to the differentiated neural cells and wherein the model is then capable of a TEER of greater than 100 Ohm×cm 2 for a period of at least 72 hours.