title: In vitro fabrication of autologous living tissue-engineered vascular grafts based on prenatally harvested ovine amniotic fluid-derived stem cells.
abstract: Amniotic fluid cells (AFCs) have been proposed as a valuable source for tissue engineering and regenerative medicine. However, before clinical implementation, rigorous evaluation of this cell source in clinically relevant animal models accepted by regulatory authorities is indispensable. Today, the ovine model represents one of the most accepted preclinical animal models, in particular for cardiovascular applications. Here, we investigate the isolation and use of autologous ovine AFCs as cell source for cardiovascular tissue engineering applications. Fetal fluids were aspirated in vivo from pregnant ewes (n = 9) and from explanted uteri post mortem at different gestational ages (n = 91). Amniotic non-allantoic fluid nature was evaluated biochemically and in vivo samples were compared with post mortem reference samples. Isolated cells revealed an immunohistochemical phenotype similar to ovine bone marrow-derived mesenchymal stem cells (MSCs) and showed expression of stem cell factors described for embryonic stem cells, such as NANOG and STAT-3. Isolated ovine amniotic fluid-derived MSCs were screened for numeric chromosomal aberrations and successfully differentiated into several mesodermal phenotypes. Myofibroblastic ovine AFC lineages were then successfully used for the in vitro fabrication of small- and large-diameter tissue-engineered vascular grafts (n = 10) and cardiovascular patches (n = 34), laying the foundation for the use of this relevant pre-clinical in vivo assessment model for future amniotic fluid cell-based therapeutic applications.
authors.affiliation: Swiss Centre for Regenerative Medicine, University of Zurich, Zurich, Switzerland.|Clinic for Cardiovascular Surgery and Department of Surgical Research, University Hospital of Zurich, Zurich, Switzerland.
authors.forename: Benedikt
authors.initials: B
authors.lastname: Weber
authors.affiliation: Swiss Centre for Regenerative Medicine, University of Zurich, Zurich, Switzerland.|Clinic for Cardiovascular Surgery and Department of Surgical Research, University Hospital of Zurich, Zurich, Switzerland.
authors.forename: Debora
authors.initials: D
authors.lastname: Kehl
authors.affiliation: Clinic of Reproductive Medicine, Department of Food Animals, Vetsuisse-Faculty University of Zurich, Zurich, Switzerland.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Bleul
authors.affiliation: IMM Recherche, Institute Mutualiste Montsouris, Paris, France.
authors.forename: Luc
authors.initials: L
authors.lastname: Behr
authors.affiliation: IMM Recherche, Institute Mutualiste Montsouris, Paris, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Sammut
authors.affiliation: Swiss Centre for Regenerative Medicine, University of Zurich, Zurich, Switzerland.|Clinic for Cardiovascular Surgery and Department of Surgical Research, University Hospital of Zurich, Zurich, Switzerland.
authors.forename: Laura
authors.initials: L
authors.lastname: Frese
authors.affiliation: Swiss Centre for Regenerative Medicine, University of Zurich, Zurich, Switzerland.|Clinic for Cardiovascular Surgery and Department of Surgical Research, University Hospital of Zurich, Zurich, Switzerland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Ksiazek
authors.affiliation: Human Genetics Laboratory, Genetica AG, Zurich, Switzerland.
authors.forename: Josef
authors.initials: J
authors.lastname: Achermann
authors.affiliation: Breeding Biology Group, Swiss Federal Institute of Technology, Zurich, Switzerland.
authors.forename: Gerald
authors.initials: G
authors.lastname: Stranzinger
authors.affiliation: Swiss Centre for Regenerative Medicine, University of Zurich, Zurich, Switzerland.|Clinic for Cardiovascular Surgery and Department of Surgical Research, University Hospital of Zurich, Zurich, Switzerland.|Institute of Clinical Chemistry, University Hospital Zurich, Zurich, Switzerland.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Robert
authors.affiliation: Department of Biomedical Engineering, Soft Tissue Biomechanics and Tissue Engineering, Eindhoven University of Technology, the Netherlands.
authors.forename: Bart
authors.initials: B
authors.lastname: Sanders
authors.affiliation: Musculo-sceletal Research Unit, Vetsuisse-Faculty, University of Zurich, Zurich, Switzerland.
authors.forename: Michele
authors.initials: M
authors.lastname: Sidler
authors.affiliation: Swiss Centre for Regenerative Medicine, University of Zurich, Zurich, Switzerland.
authors.forename: Chad E
authors.initials: CE
authors.lastname: Brokopp
authors.affiliation: Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology, ETH Zurich, Switzerland.
authors.forename: Steven T
authors.initials: ST
authors.lastname: Proulx
authors.affiliation: Department of Diagnostic Radiology, University Hospital Zurich, Zurich, Switzerland.
authors.forename: Thomas
authors.initials: T
authors.lastname: Frauenfelder
authors.affiliation: Swiss Centre for Regenerative Medicine, University of Zurich, Zurich, Switzerland.|Clinic for Cardiovascular Surgery and Department of Surgical Research, University Hospital of Zurich, Zurich, Switzerland.
authors.forename: Roman
authors.initials: R
authors.lastname: Schoenauer
authors.affiliation: Swiss Centre for Regenerative Medicine, University of Zurich, Zurich, Switzerland.|Clinic for Cardiovascular Surgery and Department of Surgical Research, University Hospital of Zurich, Zurich, Switzerland.
authors.forename: Maximilian Y
authors.initials: MY
authors.lastname: Emmert
authors.affiliation: Swiss Centre for Regenerative Medicine, University of Zurich, Zurich, Switzerland.|Clinic for Cardiovascular Surgery and Department of Surgical Research, University Hospital of Zurich, Zurich, Switzerland.
authors.forename: Volkmar
authors.initials: V
authors.lastname: Falk
authors.affiliation: Swiss Centre for Regenerative Medicine, University of Zurich, Zurich, Switzerland.|Clinic for Cardiovascular Surgery and Department of Surgical Research, University Hospital of Zurich, Zurich, Switzerland.|Centre for Applied Biotechnology and Molecular Medicine (CABMM), Zurich, Switzerland.
authors.forename: Simon P
authors.initials: SP
authors.lastname: Hoerstrup
mesh_terms: D000071916:AC133 Antigen; D000650:Amnion / Q000166:cytology*; D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001807:Blood Vessel Prosthesis; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation; D048430:Cell Shape; D002470:Cell Survival; D002869:Chromosome Aberrations; D042783:Endothelial Cells / Q000166:cytology; D005260:Female; D005838:Genotype; D006023:Glycoproteins / Q000378:metabolism; D007621:Karyotyping; D059630:Mesenchymal Stem Cells; D058628:Myofibroblasts / Q000166:cytology; D010455:Peptides / Q000378:metabolism; D010641:Phenotype; D011247:Pregnancy; D012756:Sheep; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry; D014182:Transplantation, Autologous
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; D010455:Peptides
keywords: amniotic fluid; cardiovascular tissue engineering; fetal model; mesenchymal stem cells; ovine in-vivo model; vascular graft