title: Fibroblasts derived from human pluripotent stem cells activate angiogenic responses in vitro and in vivo.
abstract: Human embryonic and induced pluripotent stem cells (hESC/hiPSC) are promising cell sources for the derivation of large numbers of specific cell types for tissue engineering and cell therapy applications. We have describe a directed differentiation protocol that generates fibroblasts from both hESC and hiPSC (EDK/iPDK) that support the repair and regeneration of epithelial tissue in engineered, 3D skin equivalents. In the current study, we analyzed the secretory profiles of EDK and iPDK cells to investigate the production of factors that activate and promote angiogenesis. Analysis of in vitro secretion profiles from EDK and iPDK cells demonstrated the elevated secretion of pro-angiogenic soluble mediators, including VEGF, HGF, IL-8, PDGF-AA, and Ang-1, that stimulated endothelial cell sprouting in a 3D model of angiogenesis in vitro. Phenotypic analysis of EDK and iPDK cells during the course of differentiation from hESCs and iPSCs revealed that both cell types progressively acquired pericyte lineage markers NG2, PDGFRβ, CD105, and CD73 and demonstrated transient induction of pericyte progenitor markers CD31, CD34, and Flk1/VEGFR2. Furthermore, when co-cultured with endothelial cells in 3D fibrin-based constructs, EDK and iPDK cells promoted self-assembly of vascular networks and vascular basement membrane deposition. Finally, transplantation of EDK cells into mice with hindlimb ischemia significantly reduced tissue necrosis and improved blood perfusion, demonstrating the potential of these cells to stimulate angiogenic responses in vivo. These findings demonstrate that stable populations of pericyte-like angiogenic cells can be generated with high efficiency from hESC and hiPSC using a directed differentiation approach. This provides new cell sources and opportunities for vascular tissue engineering and for the development of novel strategies in regenerative medicine.
authors.affiliation: Program in Cell, Molecular and Developmental Biology, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, Massachusetts, United States of America.
authors.forename: Yulia
authors.initials: Y
authors.lastname: Shamis
authors.affiliation: School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts, United States of America ; Wyss Institute For Biological Inspired Engineering, Harvard University, Boston, Massachusetts, United States of America.
authors.forename: Eduardo A
authors.initials: EA
authors.lastname: Silva
authors.affiliation: Program in Cell, Molecular and Developmental Biology, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, Massachusetts, United States of America.
authors.forename: Kyle J
authors.initials: KJ
authors.lastname: Hewitt
authors.affiliation: School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts, United States of America ; Wyss Institute For Biological Inspired Engineering, Harvard University, Boston, Massachusetts, United States of America.
authors.forename: Yevgeny
authors.initials: Y
authors.lastname: Brudno
authors.affiliation: Department of Biomedical Engineering, Technion - Israel Institute of Technology, Haifa, Israel.
authors.forename: Shulamit
authors.initials: S
authors.lastname: Levenberg
authors.affiliation: School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts, United States of America ; Department of Biomedical Engineering, Technion - Israel Institute of Technology, Haifa, Israel.
authors.forename: David J
authors.initials: DJ
authors.lastname: Mooney
authors.affiliation: Program in Cell, Molecular and Developmental Biology, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, Massachusetts, United States of America.
authors.forename: Jonathan A
authors.initials: JA
authors.lastname: Garlick
mesh_terms: D043925:Angiogenesis Inducing Agents / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D005121:Extremities / Q000098:blood supply / Q000473:pathology; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D007511:Ischemia / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D051379:Mice; D018919:Neovascularization, Physiologic; D020286:Pericytes / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D020543:Proteome; D033581:Stem Cell Transplantation
chemical_list: D043925:Angiogenesis Inducing Agents; D015415:Biomarkers; D020543:Proteome