title: c-Kit+ progenitors generate vascular cells for tissue-engineered grafts through modulation of the Wnt/Klf4 pathway.
abstract: The development of decellularised scaffolds for small diameter vascular grafts is hampered by their limited patency, due to the lack of luminal cell coverage by endothelial cells (EC) and to the low tone of the vessel due to absence of a contractile smooth muscle cells (SMC). In this study, we identify a population of vascular progenitor c-Kit+/Sca-1- cells available in large numbers and derived from immuno-privileged embryonic stem cells (ESCs). We also define an efficient and controlled differentiation protocol yielding fully to differentiated ECs and SMCs in sufficient numbers to allow the repopulation of a tissue engineered vascular graft. When seeded ex vivo on a decellularised vessel, c-Kit+/Sca-1-derived cells recapitulated the native vessel structure and upon in vivo implantation in the mouse, markedly reduced neointima formation and mortality, restoring functional vascularisation. We showed that Krüppel-like transcription factor 4 (Klf4) regulates the choice of differentiation pathway of these cells through β-catenin activation and was itself regulated by the canonical Wnt pathway activator lithium chloride. Our data show that ESC-derived c-Kit+/Sca-1-cells can be differentiated through a Klf4/β-catenin dependent pathway and are a suitable source of vascular progenitors for the creation of superior tissue-engineered vessels from decellularised scaffolds.
authors.affiliation: Department of Materials, Department of Bioengineering and Institute of Biomedical Engineering, Imperial College London, United Kingdom. Electronic address: p.campagnolo@imperial.ac.uk.
authors.forename: Paola
authors.initials: P
authors.lastname: Campagnolo
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre, King's College London, London, United Kingdom.
authors.forename: Tsung-Neng
authors.initials: TN
authors.lastname: Tsai
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre, King's College London, London, United Kingdom.
authors.forename: Xuechong
authors.initials: X
authors.lastname: Hong
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre, King's College London, London, United Kingdom.
authors.forename: John Paul
authors.initials: JP
authors.lastname: Kirton
authors.affiliation: Institute of Psychiatry, Psychology & Neuroscience, King's College London, London, United Kingdom.
authors.forename: Po-Wah
authors.initials: PW
authors.lastname: So
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre, King's College London, London, United Kingdom.
authors.forename: Andriana
authors.initials: A
authors.lastname: Margariti
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre, King's College London, London, United Kingdom.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Di Bernardini
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre, King's College London, London, United Kingdom.
authors.forename: Mei Mei
authors.initials: MM
authors.lastname: Wong
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre, King's College London, London, United Kingdom.
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Department of Materials, Department of Bioengineering and Institute of Biomedical Engineering, Imperial College London, United Kingdom.
authors.forename: Molly M
authors.initials: MM
authors.lastname: Stevens
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre, King's College London, London, United Kingdom. Electronic address: qingbo.xu@kcl.ac.uk.
authors.forename: Qingbo
authors.initials: Q
authors.lastname: Xu
mesh_terms: D000818:Animals; D000950:Antigens, Ly / Q000032:analysis / Q000378:metabolism; D001807:Blood Vessel Prosthesis; D002454:Cell Differentiation; D002460:Cell Line; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000378:metabolism*; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism; D051379:Mice; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism; D019009:Proto-Oncogene Proteins c-kit / Q000032:analysis / Q000378:metabolism*; D023822:Tissue Engineering / Q000379:methods*; D060449:Wnt Signaling Pathway
chemical_list: D000950:Antigens, Ly; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D019009:Proto-Oncogene Proteins c-kit
keywords: Cell signalling; Endothelialization; Stem cells; Vascular graft