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Title: Efficient differentiation of human embryonic stem cells to arterial and venous endothelial cells under feeder- and serum-free conditions
Authors: Sriram, G 
Tan, J.Y 
Islam, I 
Rufaihah, A.J 
Cao, T 
Keywords: bone morphogenetic protein 4
CD31 antigen
CD34 antigen
fibroblast growth factor 2
glycogen synthase kinase 3
6 [[2 [[4 (2,4 dichlorophenyl) 5 (4 methyl 1h imidazol 2 yl) 2 pyrimidinyl]amino]ethyl]amino]nicotinonitrile
bone morphogenetic protein 4
CD31 antigen
CD34 antigen
fibroblast growth factor 2
pyridine derivative
pyrimidine derivative
vasculotropin A
animal experiment
animal model
animal tissue
cell differentiation
cell function
cell migration
cell population
controlled study
culture medium
embryonic stem cell
endothelial progenitor cell
endothelium cell
human cell
in vitro study
in vivo study
priority journal
risk assessment
antagonists and inhibitors
cell differentiation
cell lineage
drug effects
endothelium cell
human embryonic stem cell
nonobese diabetic mouse
SCID mouse
vascular endothelium
Antigens, CD31
Antigens, CD34
Bone Morphogenetic Protein 4
Cell Differentiation
Cell Lineage
Endothelial Cells
Endothelium, Vascular
Fibroblast Growth Factor 2
Glycogen Synthase Kinase 3
Human Embryonic Stem Cells
Mice, Inbred NOD
Mice, SCID
Transplantation, Heterologous
Vascular Endothelial Growth Factor A
Issue Date: 2015
Citation: Sriram, G, Tan, J.Y, Islam, I, Rufaihah, A.J, Cao, T (2015). Efficient differentiation of human embryonic stem cells to arterial and venous endothelial cells under feeder- and serum-free conditions. Stem Cell Research and Therapy 6 (1) : 261. ScholarBank@NUS Repository.
Abstract: Background: Heterogeneity of endothelial cells (ECs) is a hallmark of the vascular system which may impact the development and management of vascular disorders. Despite the tremendous progress in differentiation of human embryonic stem cells (hESCs) towards endothelial lineage, differentiation into arterial and venous endothelial phenotypes remains elusive. Additionally, current differentiation strategies are hampered by inefficiency, lack of reproducibility, and use of animal-derived products. Methods: To direct the differentiation of hESCs to endothelial subtypes, H1- and H9-hESCs were seeded on human plasma fibronectin and differentiated under chemically defined conditions by sequential modulation of glycogen synthase kinase-3 (GSK-3), basic fibroblast growth factor (bFGF), bone morphogenetic protein 4 (BMP4) and vascular endothelial growth factor (VEGF) signaling pathways for 5 days. Following the initial differentiation, the endothelial progenitor cells (CD34+CD31+ cells) were sorted and terminally differentiated under serum-free conditions to arterial and venous ECs. The transcriptome and secretome profiles of the two distinct populations of hESC-derived arterial and venous ECs were characterized. Furthermore, the safety and functionality of these cells upon in vivo transplantation were characterized. Results: Sequential modulation of hESCs with GSK-3 inhibitor, bFGF, BMP4 and VEGF resulted in stages reminiscent of primitive streak, early mesoderm/lateral plate mesoderm, and endothelial progenitors under feeder- and serum-free conditions. Furthermore, these endothelial progenitors demonstrated differentiation potential to almost pure populations of arterial and venous endothelial phenotypes under serum-free conditions. Specifically, the endothelial progenitors differentiated to venous ECs in the absence of VEGF, and to arterial phenotype under low concentrations of VEGF. Additionally, these hESC-derived arterial and venous ECs showed distinct molecular and functional profiles in vitro. Furthermore, these hESC-derived arterial and venous ECs were nontumorigenic and were functional in terms of forming perfused microvascular channels upon subcutaneous implantation in the mouse. Conclusions: We report a simple, rapid, and efficient protocol for directed differentiation of hESCs into endothelial progenitor cells capable of differentiation to arterial and venous ECs under feeder-free and serum-free conditions. This could offer a human platform to study arterial-venous specification for various applications related to drug discovery, disease modeling and regenerative medicine in the future. © 2015 Sriram et al.
Source Title: Stem Cell Research and Therapy
ISSN: 17576512
DOI: 10.1186/s13287-015-0260-5
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