Inflammatory priming enhances mesenchymal stromal cell secretome potential as a clinical product for regenerative medicine approaches through secreted factors and EV-miRNAs: the example of joint disease

摘要: Mesenchymal stromal cell (MSC)-enriched products showed positive clinical outcomes in regenerative medicine, where tissue restoration and inflammation control are needed. GMP-expanded MSCs displayed an even higher potential due to exclusive secretion of therapeutic factors, both free conveyed within extracellular vesicles (EVs), collectively termed secretome. Moreover, priming with biochemical cues may influence the portfolio biological activities MSC-derived factors. For these reasons, use naive or primed secretome gained attention as a cell-free option. Albeit, at present, homogenous comprehensive fingerprint is still missing. Therefore, aim this work was deeply characterize adipose-derived MSC (ASC)-secreted factors EV-miRNAs, their modulation after IFNγ preconditioning. The crucial target pathology type also scored osteoarthritis evaluate disease-driven potency. ASCs were isolated from four donors cultured without IFNγ. Two-hundred secreted assayed by ELISA. ASC-EVs ultracentrifugation validated flow cytometry, transmission electron microscopy, nanoparticle tracking analysis. miRNome deciphered high-throughput screening. Bioinformatics used predict modulatory effect molecules on pathologic cartilage synovial macrophages based public datasets. Models for chondrocytes test cytometry anti-inflammatory Data that more than 60 cytokines/chemokines could be identified varying levels intensity all samples. vast majority involved matrix remodeling, chemotaxis motility inflammatory cells. able further increase capacity stimulate migration signals. 240 miRNAs found ASC-EVs. Sixty accounted > 95% genetic message resulted chondro-protective M2 macrophage polarizing. Inflammation tipped balance towards pronounced phenotype. In silico data confirmed inflamed chondrocytes, being phenotype marker CD163 reduce chondrocyte VCAM1, respectively. enhanced Given soluble ASC marked modulate degenerative processes. Preconditioning ability, suggesting effective strategy obtain potent product which should always driven molecular mark pathology.