Abstract
Human pluripotent stem cell (hPSC)-derived endothelial cells and their progenitors may provide the means for vascularization of tissue-engineered constructs and can serve as models to study vascular development and disease. Here, we report a method to efficiently produce endothelial cells from hPSCs via GSK3 inhibition and culture in defined media to direct hPSC differentiation to CD34(+)CD31(+) endothelial progenitors. Exogenous vascular endothelial growth factor (VEGF) treatment was dispensable, and endothelial progenitor differentiation was β-catenin dependent. Furthermore, by clonal analysis, we showed that CD34(+)CD31(+)CD117(+)TIE-2(+) endothelial progenitors were multipotent, capable of differentiating into calponin-expressing smooth muscle cells and CD31(+)CD144(+)vWF(+)I-CAM1(+) endothelial cells. These endothelial cells were capable of 20 population doublings, formed tube-like structures, imported acetylated low-density lipoprotein, and maintained a dynamic barrier function. This study provides a rapid and efficient method for production of hPSC-derived endothelial progenitors and endothelial cells and identifies WNT/β-catenin signaling as a primary regulator for generating vascular cells from hPSCs.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Aminopyridines / pharmacology
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Antigens, CD34 / genetics
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Antigens, CD34 / metabolism
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Blotting, Western
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Cell Differentiation*
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Cells, Cultured
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Embryonic Stem Cells / cytology
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Embryonic Stem Cells / metabolism
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Endothelial Cells / cytology
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Endothelial Cells / metabolism
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Endothelial Progenitor Cells / cytology*
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Endothelial Progenitor Cells / metabolism
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Flow Cytometry
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Gene Expression
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Glycogen Synthase Kinase 3 / antagonists & inhibitors
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Glycogen Synthase Kinase 3 / metabolism
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Humans
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Induced Pluripotent Stem Cells / cytology
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Induced Pluripotent Stem Cells / metabolism
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Platelet Endothelial Cell Adhesion Molecule-1 / genetics
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Platelet Endothelial Cell Adhesion Molecule-1 / metabolism
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Pluripotent Stem Cells / cytology*
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Pluripotent Stem Cells / metabolism
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Pyrimidines / pharmacology
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Reverse Transcriptase Polymerase Chain Reaction
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Vascular Endothelial Growth Factor A / pharmacology
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Wnt Signaling Pathway*
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beta Catenin / genetics
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beta Catenin / metabolism
Substances
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Aminopyridines
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Antigens, CD34
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Chir 98014
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Platelet Endothelial Cell Adhesion Molecule-1
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Pyrimidines
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Vascular Endothelial Growth Factor A
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beta Catenin
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Glycogen Synthase Kinase 3