Priming equine bone marrow derived mesenchymal stem cells (eBM-MSCs) with pro-inflammatory cytokines: implications in immunomodulation-immunogenicity balance, cell viability and differentiation potential

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 TNFa and IFN¿ 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

Scaffolds descelularizados derivados de matriz extracelular dérmica humana: caracterizaciones composicionales, mecánicas y biológicas in vitro

Currently, there is a growing demand for in vitro skin models. In search of a more realistic approximation of the tissue microenvironment, here, we present an optimized protocol for extracting and characterizing hydrogels obtained from decellularized dermal extracellular matrix from human skin to generate scaffolds for subsequent use in 3D skin models.Actualmente, existe una creciente demanda de modelos de piel in vitro. En busca de una aproximación más realista del microambiente del tejido, aquí presentamos un protocolo optimizado para extraer y caracterizar hidrogeles obtenidos a partir de la matriz extracelular dérmica decelularizada de piel humana, con el fin de generar andamios para su posterior uso en modelos de piel en 3D