title: Defining minimum essential factors to derive highly pure human endothelial cells from iPS/ES cells in an animal substance-free system.
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.
authors: Institute of Clinical Medicine, National Cheng Kung University, Tainan, Taiwan, ROC.
authors: Yu-Ting
authors: YT
authors: Wu
authors: Laboratory Animal Center, College of Medicine, National Taiwan University, Taipei, Taiwan, ROC.
authors: I-Shing Yu
authors: Institute of Clinical Medicine, National Cheng Kung University, Tainan, Taiwan, ROC.
authors: Kuen-Jer
authors: KJ
authors: Tsai
authors: Institute of Clinical Medicine, National Cheng Kung University, Tainan, Taiwan, ROC.
authors: Chien-Yu
authors: CY
authors: Shih
authors: Food Industry Research and Development Institute, Hsinchu, Taiwan, ROC.
authors: Shiaw-Min
authors: SM
authors: Hwang
authors: Division of Infectious Diseases, National Health Research Institutes, Tainan, Taiwan, ROC.
authors: Ih-Jen
authors: IJ
authors: Su
authors: Institute of Clinical Medicine, National Cheng Kung University, Tainan, Taiwan, ROC.
authors: Po-Min
authors: PM
authors: Chiang
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D038341:Glycogen Synthase Kinases / Q000037:antagonists & inhibitors; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D008648:Mesoderm / Q000196:embryology / Q000378:metabolism; D051379:Mice; D016212:Transforming Growth Factor beta / Q000819:agonists; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism; D042442:Vascular Endothelial Growth Factors / Q000378:metabolism; D051153:Wnt Proteins / Q000819:agonists
chemical_list: D016212:Transforming Growth Factor beta; D042442:Vascular Endothelial Growth Factors; D051153:Wnt Proteins; D016222:Fibroblast Growth Factor 2; D040301:Vascular Endothelial Growth Factor Receptor-2; D038341:Glycogen Synthase Kinases