VEGF Induces Differentiation of Functional Endothelium From Human Embryonic Stem Cells
Open Access
- 1 January 2010
- journal article
- research article
- Published by Ovid Technologies (Wolters Kluwer Health) in Arteriosclerosis, Thrombosis, and Vascular Biology
- Vol. 30 (1), 80-89
- https://doi.org/10.1161/atvbaha.109.194233
Abstract
Objective— Human embryonic stem cells (hESCs) offer a sustainable source of endothelial cells for therapeutic vascularization and tissue engineering, but current techniques for generating these cells remain inefficient. We endeavored to induce and isolate functional endothelial cells from differentiating hESCs. Methods and Results— To enhance endothelial cell differentiation above a baseline of ≈2% in embryoid body (EB) spontaneous differentiation, 3 alternate culture conditions were compared. Vascular endothelial growth factor (VEGF) treatment of EBs showed the best induction, with markedly increased expression of endothelial cell proteins CD31, VE-Cadherin, and von Willebrand Factor, but not the hematopoietic cell marker CD45. CD31 expression peaked around days 10 to 14. Continuous VEGF treatment resulted in a 4- to 5-fold enrichment of CD31 + cells but did not increase endothelial proliferation rates, suggesting a primary effect on differentiation. CD31 + cells purified from differentiating EBs upregulated ICAM-1 and VCAM-1 in response to TNFα, confirming their ability to function as endothelial cells. These cells also expressed multiple endothelial genes and formed lumenized vessels when seeded onto porous poly(2-hydroxyethyl methacrylate) scaffolds and implanted in vivo subcutaneously in athymic rats. Collagen gel constructs containing hESC-derived endothelial cells and implanted into infarcted nude rat hearts formed robust networks of patent vessels filled with host blood cells. Conclusions— VEGF induces functional endothelial cells from hESCs independent of endothelial cell proliferation. This enrichment method increases endothelial cell yield, enabling applications for revascularization as well as basic studies of human endothelial biology. We demonstrate the ability of hESC-derived endothelial cells to facilitate vascularization of tissue-engineered implants.Keywords
This publication has 39 references indexed in Scilit:
- Augmentation of Neovascularizaiton in Hindlimb Ischemia by Combined Transplantation of Human Embryonic Stem Cells-Derived Endothelial and Mural CellsPLOS ONE, 2008
- Vascular Progenitor Cells Isolated From Human Embryonic Stem Cells Give Rise to Endothelial and Smooth Muscle–Like Cells and Form Vascular Networks In VivoCirculation Research, 2007
- Photo-patterning of porous hydrogels for tissue engineeringBiomaterials, 2007
- Biphasic role for Wnt/β-catenin signaling in cardiac specification in zebrafish and embryonic stem cellsProceedings of the National Academy of Sciences of the United States of America, 2007
- Endothelial cells derived from human embryonic stem cells form durable blood vessels in vivoNature Biotechnology, 2007
- Development of the hemangioblast defines the onset of hematopoiesis in human ES cell differentiation culturesBlood, 2006
- Redefining endothelial progenitor cells via clonal analysis and hematopoietic stem/progenitor cell principalsBlood, 2006
- VEGF receptor signalling ? in control of vascular functionNature Reviews Molecular Cell Biology, 2006
- A hierarchical order of factors in the generation of FLK1- and SCL-expressing hematopoietic and endothelial progenitors from embryonic stem cellsDevelopment, 2004
- Abnormal blood vessel development and lethality in embryos lacking a single VEGF alleleNature, 1996