title: Perivascular Mesenchymal Stem Cells in Sheep: Characterization and Autologous Transplantation in a Model of Articular Cartilage Repair.
abstract: Previous research has indicated that purified perivascular stem cells (PSCs) have increased chondrogenic potential compared to conventional mesenchymal stem cells (MSCs) derived in culture. This study aimed to develop an autologous large animal model for PSC transplantation and to specifically determine if implanted cells are retained in articular cartilage defects. Immunohistochemistry and fluorescence-activated cell sorting were used to ascertain the reactivity of anti-human and anti-ovine antibodies, which were combined and used to identify and isolate pericytes (CD34-CD45-CD146+) and adventitial cells (CD34+CD45-CD146-). The purified cells demonstrated osteogenic, adipogenic, and chondrogenic potential in culture. Autologous ovine PSCs (oPSCs) were isolated, cultured, and efficiently transfected using a green fluorescence protein (GFP) encoding lentivirus. The cells were implanted into articular cartilage defects on the medial femoral condyle using hydrogel and collagen membranes. Four weeks following implantation, the condyle was explanted and confocal laser scanning microscopy demonstrated the presence of oPSCs in the defect repaired with the hydrogel. These data suggest the testability in a large animal of native MSC autologous grafting, thus avoiding possible biases associated with xenotransplantation. Such a setting will be used in priority for indications in orthopedics, at first to model articular cartilage repair.
authors.affiliation: 1 MRC Centre for Regenerative Medicine, The University of Edinburgh , Edinburgh, United Kingdom .|2 Department of Trauma and Orthopaedic Surgery, The Royal Infirmary of Edinburgh , Edinburgh, United Kingdom .
authors.forename: Paul
authors.initials: P
authors.lastname: Hindle
authors.affiliation: 1 MRC Centre for Regenerative Medicine, The University of Edinburgh , Edinburgh, United Kingdom .
authors.forename: James
authors.initials: J
authors.lastname: Baily
authors.affiliation: 1 MRC Centre for Regenerative Medicine, The University of Edinburgh , Edinburgh, United Kingdom .
authors.forename: Nusrat
authors.initials: N
authors.lastname: Khan
authors.affiliation: 2 Department of Trauma and Orthopaedic Surgery, The Royal Infirmary of Edinburgh , Edinburgh, United Kingdom .
authors.forename: Leela C
authors.initials: LC
authors.lastname: Biant
authors.affiliation: 2 Department of Trauma and Orthopaedic Surgery, The Royal Infirmary of Edinburgh , Edinburgh, United Kingdom .
authors.forename: A Hamish R
authors.initials: AH
authors.lastname: Simpson
authors.affiliation: 1 MRC Centre for Regenerative Medicine, The University of Edinburgh , Edinburgh, United Kingdom .|3 The University of California , Los Angeles, Los Angeles, California.
authors.forename: Bruno
authors.initials: B
authors.lastname: Péault
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D001808:Blood Vessels / Q000166:cytology*; D002358:Cartilage, Articular / Q000187:drug effects / Q000473:pathology*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003429:Cross Reactions / Q000276:immunology; D004195:Disease Models, Animal; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000378:metabolism; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000494:pharmacology; D007150:Immunohistochemistry; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D020286:Pericytes / Q000166:cytology / Q000187:drug effects; D012756:Sheep; D014162:Transfection; D014182:Transplantation, Autologous; D014945:Wound Healing / Q000187:drug effects
chemical_list: D000906:Antibodies; D049452:Green Fluorescent Proteins; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate
keywords: CD34+; autologous stem cell transplantation; green fluorescent protein; in vivo tracking; pericytes; sheep model