title: Priming Equine Bone Marrow-Derived Mesenchymal Stem Cells with Proinflammatory Cytokines: Implications in Immunomodulation-Immunogenicity Balance, Cell Viability, and Differentiation Potential.
abstract: Mesenchymal stem cells (MSCs) have a great potential for treating equine musculoskeletal injuries. Although their mechanisms of action are not completely known, their immunomodulatory properties appear to be key in their functions. The expression of immunoregulatory molecules by MSCs is regulated by proinflammatory cytokines; so inflammatory priming of MSCs might improve their therapeutic potential. However, inflammatory environment could also increase MSC immunogenicity and decrease MSC viability and differentiation capacity. The aim of this study was to assess the effect of cytokine priming on equine bone marrow-derived MSC (eBM-MSC) immunoregulation, immunogenicity, viability, and differentiation potential, to enhance MSC immunoregulatory properties, without impairing their immune-evasive status, viability, and plasticity. Equine BM-MSCs (n = 4) were exposed to 5 ng/mL of TNFα and IFNγ for 12 h (CK5-priming). Subsequently, expression of genes coding for immunomodulatory, immunogenic, and apoptosis-related molecules was analyzed by real-time quantitative polymerase chain reaction. Chromatin integrity and proliferation assays were assessed to evaluate cell viability. Trilineage differentiation was evaluated by specific staining and gene expression. Cells were reseeded in a basal medium for additional 7 days post-CK5 to elucidate if priming-induced changes were maintained along the time. CK5-priming led to an upregulation of immunoregulatory genes IDO, iNOS, IL-6, COX-2, and VCAM-1. MHC-II and CD40 were also upregulated, but no change in other costimulatory molecules was observed. These changes were not maintained 7 days after CK5-priming. Viability and differentiation potential were maintained after CK5-priming. These findings suggest that CK5-priming of eBM-MSCs could improve their in vivo effectiveness without affecting other eBM-MSC properties.
authors.affiliation: 1 Laboratorio de Genetica Bioquimica LAGENBIO (Universidad de Zaragoza), Instituto Agroalimentario de Aragon- IA2 - (Universidad de Zaragoza-CITA), Instituto de Investigacion Sanitaria de Aragon (IIS) , Zaragoza, Spain .|2 Servicio de Cirugía y Medicina Equina, Hospital Veterinario, Universidad de Zaragoza , Zaragoza, Spain .
authors.forename: Laura
authors.initials: L
authors.lastname: Barrachina
authors.affiliation: 1 Laboratorio de Genetica Bioquimica LAGENBIO (Universidad de Zaragoza), Instituto Agroalimentario de Aragon- IA2 - (Universidad de Zaragoza-CITA), Instituto de Investigacion Sanitaria de Aragon (IIS) , Zaragoza, Spain .
authors.forename: Ana Rosa
authors.initials: AR
authors.lastname: Remacha
authors.affiliation: 1 Laboratorio de Genetica Bioquimica LAGENBIO (Universidad de Zaragoza), Instituto Agroalimentario de Aragon- IA2 - (Universidad de Zaragoza-CITA), Instituto de Investigacion Sanitaria de Aragon (IIS) , Zaragoza, Spain .|2 Servicio de Cirugía y Medicina Equina, Hospital Veterinario, Universidad de Zaragoza , Zaragoza, Spain .
authors.forename: Antonio
authors.initials: A
authors.lastname: Romero
authors.affiliation: 1 Laboratorio de Genetica Bioquimica LAGENBIO (Universidad de Zaragoza), Instituto Agroalimentario de Aragon- IA2 - (Universidad de Zaragoza-CITA), Instituto de Investigacion Sanitaria de Aragon (IIS) , Zaragoza, Spain .|2 Servicio de Cirugía y Medicina Equina, Hospital Veterinario, Universidad de Zaragoza , Zaragoza, Spain .
authors.forename: Francisco José
authors.initials: FJ
authors.lastname: Vázquez
authors.affiliation: 1 Laboratorio de Genetica Bioquimica LAGENBIO (Universidad de Zaragoza), Instituto Agroalimentario de Aragon- IA2 - (Universidad de Zaragoza-CITA), Instituto de Investigacion Sanitaria de Aragon (IIS) , Zaragoza, Spain .|3 Servicio de Cirugía Ortopédica y Traumatología, Hospital Clínico Universitario Lozano Blesa , Zaragoza, Zaragoza, Spain .
authors.forename: Jorge
authors.initials: J
authors.lastname: Albareda
authors.affiliation: 1 Laboratorio de Genetica Bioquimica LAGENBIO (Universidad de Zaragoza), Instituto Agroalimentario de Aragon- IA2 - (Universidad de Zaragoza-CITA), Instituto de Investigacion Sanitaria de Aragon (IIS) , Zaragoza, Spain .|4 Departament de Medicina i Cirugia Animal, Universidad Autónoma de Barcelona , Barcelona, Spain .
authors.forename: Marta
authors.initials: M
authors.lastname: Prades
authors.affiliation: 5 Departamento de Biología, Universidad Autónoma de Madrid, Ciudad Universitaria de Cantoblanco , Madrid, Spain .
authors.forename: Jaime
authors.initials: J
authors.lastname: Gosálvez
authors.affiliation: 5 Departamento de Biología, Universidad Autónoma de Madrid, Ciudad Universitaria de Cantoblanco , Madrid, Spain .
authors.forename: Rosa
authors.initials: R
authors.lastname: Roy
authors.affiliation: 1 Laboratorio de Genetica Bioquimica LAGENBIO (Universidad de Zaragoza), Instituto Agroalimentario de Aragon- IA2 - (Universidad de Zaragoza-CITA), Instituto de Investigacion Sanitaria de Aragon (IIS) , Zaragoza, Spain .
authors.forename: Pilar
authors.initials: P
authors.lastname: Zaragoza
authors.affiliation: 1 Laboratorio de Genetica Bioquimica LAGENBIO (Universidad de Zaragoza), Instituto Agroalimentario de Aragon- IA2 - (Universidad de Zaragoza-CITA), Instituto de Investigacion Sanitaria de Aragon (IIS) , Zaragoza, Spain .
authors.forename: Inmaculada
authors.initials: I
authors.lastname: Martín-Burriel
authors.affiliation: 1 Laboratorio de Genetica Bioquimica LAGENBIO (Universidad de Zaragoza), Instituto Agroalimentario de Aragon- IA2 - (Universidad de Zaragoza-CITA), Instituto de Investigacion Sanitaria de Aragon (IIS) , Zaragoza, Spain .
authors.forename: Clementina
authors.initials: C
authors.lastname: Rodellar
mesh_terms: D050156:Adipogenesis / Q000187:drug effects / Q000235:genetics; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D019070:Cell Lineage / Q000187:drug effects / Q000235:genetics; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D020219:Chondrogenesis / Q000187:drug effects / Q000235:genetics; D016207:Cytokines / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D006736:Horses; D056747:Immunomodulation / Q000187:drug effects*; D018836:Inflammation Mediators / Q000494:pharmacology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics
chemical_list: D016207:Cytokines; D018836:Inflammation Mediators
keywords: horse; immune regulation; immunogenicity; inflammation; regenerative medicine