title: A reproducible method for the isolation and expansion of ovine mesenchymal stromal cells from bone marrow for use in regenerative medicine preclinical studies.
abstract: The use of multipotent mesenchymal stromal cells (MSCs) as candidate medicines for treating a variety of pathologies is based on their qualities as either progenitors for the regeneration of damaged tissue or producers of a number of molecules with pharmacological properties. Preclinical product development programmes include the use of well characterized cell populations for proof of efficacy and safety studies before testing in humans. In the field of orthopaedics, an increasing number of translational studies use sheep as an in vivo test system because of the similarities with humans in size and musculoskeletal architecture. However, robust and reproducible methods for the isolation, expansion, manipulation and characterization of ovine MSCs have not yet been standardised. The present study describes a method for isolation and expansion of fibroblastic-like, adherent ovine MSCs that express CD44, CD90, CD140a, CD105 and CD166, and display trilineage differentiation potential. The 3-week bioprocess proposed here typically yielded cell densities of 1.4 × 104MSCs/cm2at passage 2, with an expansion factor of 37.8 and approximately eight cumulative population doublings. The osteogenic potential of MSCs derived following this methodology was further evaluated in vivo in a translational model of osteonecrosis of the femoral head, in which the persistence of grafted cells in the host tissue and their lineage commitment into osteoblasts and osteocytes was demonstrated by tracking enhanced green fluorescent protein-labelled cells. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Servei de Teràpia Cel·lular, Banc de Sang i Teixits, Edifici Dr. Frederic Duran i Jordà, Barcelona, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Caminal
authors.affiliation: Orthopaedic Surgery and Traumatology, Hospital Universitari Vall d'Hebron, Barcelona, Spain.
authors.forename: Roberto
authors.initials: R
authors.lastname: Vélez
authors.affiliation: Departament de Medicina i Cirurgia Animals, Àrea de Medicina i Cirurgia Animal, Facultat de Veterinària, Universitat Autònoma de Barcelona, Cerdanyola del Vallès, Spain.
authors.forename: Rosa Maria
authors.initials: RM
authors.lastname: Rabanal
authors.affiliation: Servei de Teràpia Cel·lular, Banc de Sang i Teixits, Edifici Dr. Frederic Duran i Jordà, Barcelona, Spain.
authors.forename: Daniel
authors.initials: D
authors.lastname: Vivas
authors.affiliation: Tissue Engineering Unit, Centre for Genomic Regulation, Barcelona, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Batlle-Morera
authors.affiliation: Orthopaedic Surgery and Traumatology, Hospital Universitari Vall d'Hebron, Barcelona, Spain.
authors.forename: Màrius
authors.initials: M
authors.lastname: Aguirre
authors.affiliation: Grup de Teràpia Gènica i Cel·lular, Institut de Recerca (VHIR), Hospital Universitari Vall d'Hebron, Barcelona, Spain.
authors.forename: Jordi
authors.initials: J
authors.lastname: Barquinero
authors.affiliation: Servei de Teràpia Cel·lular, Banc de Sang i Teixits, Edifici Dr. Frederic Duran i Jordà, Barcelona, Spain.|Chair of Transfusion Medicine and Cellular and Tissue Therapies, Universitat Autònoma de Barcelona,Campus UAB, Bellaterra, Spain.
authors.forename: Joan
authors.initials: J
authors.lastname: García
authors.affiliation: Servei de Teràpia Cel·lular, Banc de Sang i Teixits, Edifici Dr. Frederic Duran i Jordà, Barcelona, Spain.
authors.forename: Joaquim
authors.identifier: 0000-0001-9719-5235
authors.initials: J
authors.lastname: Vives
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D005260:Female; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D044968:Regenerative Medicine / Q000379:methods*; D015203:Reproducibility of Results; D012756:Sheep; D023822:Tissue Engineering; D054457:Tissue Scaffolds / Q000737:chemistry
keywords: femoral head osteonecrosis; multipotent mesenchymal stromal cell; orthopaedics; sheep; tissue engineering; translational research