Retinoic Acid Enhanced Human Stem Cell Derived Blood Brain Barrier Model
First Claim
1. A method of creating a fully-human blood-brain barrier (BBB) model, comprising the steps of:
- (a) obtaining a mixture of neural cells and brain microvascular endothelial cells (BMECs), wherein the neural cells and BMECs that comprise the mixture were produced from the differentiation of human pluripotent stem cells (hPSCs);
(b) purifying BMECs from the mixture of neural cells and BMECs of step(a); and
(c) co-culturing the purified BMECs with a cell type selected from the group consisting of pericytes, astrocytes and differentiated neural progenitor cells (NPCs), wherein a blood brain barrier model is created.
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Abstract
In one embodiment, the present invention is a method of creating a fully-human blood-brain barrier (BBB) model, comprising the steps of (a) obtaining a mixture of neural cells and brain microvascular endothelial cells (BMECs), wherein the neural cells and BMECs that comprise the mixture were produced from the differentiation of human pluripotent stem cells (hPSCs); (b) purifying BMECs from the mixture of neural cells and BMECs of step (a); and (c) co-culturing the purified BMECs with a cell type selected from the group consisting of pericytes, astrocytes and differentiated neural progenitor cells (NPCs), wherein a blood brain barrier model is created.
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Citations
20 Claims
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1. A method of creating a fully-human blood-brain barrier (BBB) model, comprising the steps of:
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(a) obtaining a mixture of neural cells and brain microvascular endothelial cells (BMECs), wherein the neural cells and BMECs that comprise the mixture were produced from the differentiation of human pluripotent stem cells (hPSCs); (b) purifying BMECs from the mixture of neural cells and BMECs of step(a); and (c) co-culturing the purified BMECs with a cell type selected from the group consisting of pericytes, astrocytes and differentiated neural progenitor cells (NPCs), wherein a blood brain barrier model is created. - View Dependent Claims (2, 3, 4, 5)
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6. A method of creating a retinoic acid (RA)-enhanced or RA-like compound-enhanced mammalian blood-brain barrier (BBB) model, comprising the steps of:
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(a) obtaining a mixture of neural cells and brain microvascular endothelial cells (BMECs) in the presence of RA or RA-like compound, wherein the mixture of neural cells and BMECs was produced from the differentiation of human pluripotent stem cells (hPSCs); (b) purifying BMECs from the mixture of neural cells and BMECs; and (c) co-culturing the purified BMECs with a cell type selected from the group consisting of astrocytes, pericytes and differentiated neural progenitor cells (NPCs), wherein a BBB model is created.
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7. A mammalian blood-brain barrier (BBB) model created by the method of creating a retinoic acid (RA)-enhanced or RA-like compound-enhanced mammalian blood-brain barrier (BBB) model, the method comprising the steps of:
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a) obtaining a mixture of neural cells and brain microvascular endothelial cells (BMECs) in the presence of RA or RA-like compound, wherein the mixture of neural cells and BMECs was produced from the differentiation of human pluripotent stem cells (hPSCs); b) purifying BMECs from the mixture of neural cells and BMECs; and c) co-culturing the purified BMECs with a cell type selected from the group consisting of astrocytes, pericytes and differentiated neural progenitor cells (NPCs), wherein a BBB model is created.
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8. A method of creating a retinoic acid (RA)-enhanced or RA-like compound-enhanced fully-human blood-brain barrier (BBB) model in an optimized endothelial cell medium (OECM) wherein the OECM does not contain basic fibroblast growth factor (bFGF), comprising the steps of:
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(a) supplying a mixture of neural cells and brain microvascular endothelial cells (BMECs) in the presence of RA or RA-like compound, wherein the mixture of neural cells and BMECs was produced from the differentiation of human pluripotent stem cells (hPSCs); (b) purifying BMECs from the mixture of neural cells and BMECs; and (c) co-culturing the purified BMECs with a cell type selected from the group consisting of astrocytes, pericytes and differentiated NPCs in OECM. - View Dependent Claims (9, 10, 11, 12, 13, 14)
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15. A mammalian blood-brain barrier (BBB) model created by the method of creating a retinoic acid (RA)-enhanced or RA-like compound-enhanced fully-human blood-brain barrier (BBB) model in an optimized endothelial cell medium (OECM) wherein the OECM does not contain basic fibroblast growth factor (bFGF), the method comprising the steps of:
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a) supplying a mixture of neural cells and brain microvascular endothelial cells (BMECs) in the presence of RA or RA-like compound, wherein the mixture of neural cells and BMECs was produced from the differentiation of human pluripotent stem cells (hPSCs); b) purifying BMECs from the mixture of neural cells and BMECs; and c) co-culturing the purified BMECs with a cell type selected from the group consisting of astrocytes, pericytes and differentiated NPCs in OECM.
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16. A retinoic acid (RA) or RA-like compound-enhanced mammalian blood-brain barrier (BBB) model in optimized endothelial cell medium (OECM) expressing a TEER greater than 4000 Ohm×
- cm2, wherein the OECM does not contain basic fibroblast growth factor (bFGF), comprising;
(a) brain microvascular endothelial cells (BMECs) wherein BMECs have been purified from a mixture of neural cells and BMECs, wherein the mixture of neural cells and BMECs was produced from the differentiation of human pluripotent stem cells (hPSCs) in the presence of RA or RA-like compound; and (b) a cell type selected from the group consisting of astrocytes, pericytes and differentiated neural progenitor cells (NPCs), wherein the cell type was co-cultured with the purified BMECs in OECM such that the purified BMECs form a monolayer wherein the cells were confluent and the TEER of the confluent monolayer may be measured at greater than 4000 Ohm×
cm2. - View Dependent Claims (17, 18, 19, 20)
- cm2, wherein the OECM does not contain basic fibroblast growth factor (bFGF), comprising;
Specification