1. Defining minimum essential factors to derive highly pure human endothelial cells from iPS/ES cells in an animal substance-free system.
- Author
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Wu YT, I-Shing Yu, Tsai KJ, Shih CY, Hwang SM, Su IJ, and Chiang PM
- Subjects
- Animals, Cell Line, Embryonic Stem Cells drug effects, Endothelial Cells drug effects, Fibroblast Growth Factor 2 metabolism, Glycogen Synthase Kinases antagonists & inhibitors, Humans, Induced Pluripotent Stem Cells drug effects, Mesoderm embryology, Mesoderm metabolism, Mice, Transforming Growth Factor beta agonists, Vascular Endothelial Growth Factor Receptor-2 metabolism, Vascular Endothelial Growth Factors metabolism, Wnt Proteins agonists, Cell Differentiation drug effects, Embryonic Stem Cells cytology, Embryonic Stem Cells metabolism, Endothelial Cells cytology, Endothelial Cells metabolism, Induced Pluripotent Stem Cells cytology, Induced Pluripotent Stem Cells metabolism
- Abstract
It is desirable to obtain unlimited supplies of endothelial cells for research and therapeutics. However, current methods of deriving endothelial cells from humans suffer from issues, such as limited supplies, contamination from animal substances, and lengthy/complicated procedures. In this article we developed a way to differentiate human iPS and ES cells to highly pure endothelial cells in 5 days. The chemically defined system is robust, easy to perform, and free of animal substances. Using the system, we verified that combined TGFβ and canonical Wnt agonists are essential and sufficient for iPS/ES cell-to-mesoderm transition. Besides, VEGF-KDR signaling alone is required for endothelial formation at high density while supplementation with FGF allows for colonial endothelial differentiation. Finally, anti-adsorptive agents could enrich the endothelial output by allowing selective attachment of the endothelial precursors. The system was validated to work on multiple iPS/ES cells lines to produce endothelial cells capable of forming capillary-like structures in vitro and integrating into host vasculature in vivo. In sum, the simple yet robust differentiation system permits the unlimited supply of human endothelial cells. The defined and animal substance-free nature of the system is compatible with clinical applications and characterization of endothelial differentiation in an unbiased manner.
- Published
- 2015
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