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US 12,564,610 B2Granted
Human periventricular endothelial cell therapy for neuropsychiatric disorders
AssigneeThe McLean Hospital Corporation
InventorAnju Vasudevan
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Abstract
Described herein are methods for generating human forebrain endothelial cells, compositions comprising the cells, and methods of use thereof in therapy. Provided herein are methods for generation of human embryonic forebrain-like endothelial cells (e.g., periventricular endothelial cells) from human embryonic stem cells; the methods include addition or GABA and WNT7A for efficient differentiation, and isolation of GABRB37CD31+ cell population by FACS.
Claims14
A method of generating a population of human forebrain periventricular endothelial cells, the method comprising:
- providing a population of pluripotent stem cells:
- (i) culturing the population of pluripotent stem cells in a first, stem cell media comprising Wnt7a protein and gamma amino-butyric acid (GABA), for a first time period sufficient to generate mesodermal cells;
- (ii) culturing the mesodermal cells in a second, vascular inducing medium comprising an inhibitor of transforming growth factor beta (TGFβ) signaling, vascular endothelial growth factor-A (VEGF-A), Wnt Family Member 7A (WNT7A), and GABA, for a time sufficient to generate endothelial-like cells;
- (iii) culturing the endothelial-like cells in a third, endothelial cell culture medium, comprising growth factors and GABA, for a time sufficient for development of a population of cells comprising CD31+ gamma-aminobutyric acid type A receptor subunit beta3 (GABRB3+) endothelial cells, and
- (iv) optionally isolating CD31+GABRB3+ cells from the mixture,
- thereby generating a population of human forebrain periventricular endothelial cells.
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