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US 2017/0283772 A1Published application
Methods for differentiation of human pluripotent stem cells to brain microvascular endothelial cells
AssigneeWisconsin Alumni Research Foundation
InventorsTongcheng Qian, Eric V. Shusta, Sean P. Palecek
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Abstract
Methods for generating functional brain microvascular endothelial cells (BMECs) under chemically defined, serum-free conditions are provided. In particular, efficient and cost-effective methods for generating functional BMECs under chemically defined culture conditions are provided. BMECs obtained according to the methods provided herein are suitable for in vitro blood brain barrier (BBB) formation.
Claims10
A method for generating a population of human brain microvascular endothelial cells (BMECs) from human pluripotent stem cells, wherein the method comprises, in order,
- (a) culturing human pluripotent stem cells for about 24 hours in a chemically defined, serum-free culture medium that comprises an activator of Wnt/β-catenin signaling, whereby cells that express mesodermal markers are obtained;
- (b) culturing the cells expressing mesodermal markers for about 5 days in the presence of a chemically defined, serum-free culture medium comprising B27 supplement, whereby cells that express endothelial progenitor marker Flk-1 are obtained;
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- (c) culturing the Flk-1+ cells of (b) for about two days in the presence of a chemically defined, serum-free endothelial medium comprising B27 supplement, bFGF/FGF2, and retinoic acid (RA), whereby a cell population comprising human BMECs is obtained.
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