E = 4.4) and CSAM (detected only in CRC tissue), considering that their expression levels in plasma EVs from CRC patients had been also considerably higher than these from healthier donors in EV-ELISA Mitogen-Activated Protein Kinase 8 (MAPK8/JNK1) Proteins custom synthesis assays utilizing independent validation set. IHC staining evaluation also demonstrated four EV proteins particularly overexpressed in CRC cells. Interestingly, uptake of STAM++ EVs enhanced each proliferation and invasion of recipient cells in vitro. Conclusions:As a result TMAM, STAM, GAM and CSAM on EVs needs to be possible diagnostic or prognostic biomarkers for CRC, leading to development of precise, non-invasive and low-cost blood liquid biopsy tests in future.Saturday, May possibly 20,Area: Harbour Ballroom Symposium Session 24 EV Functions in Inflammation Chairs: Saara Laitinen and Takahiro Ochiya 1:30:00 p.m.OS24.Extracellular vesicles from adipose-derived mesenchymal stem cells market autophagy in human osteoarthritic chondrocytes Miguel Tofi -Vian1, Maria Jose Alcaraz1, Maria Dolores Perez del Caz2, Miguel Angel Castejon3 and Isabel Guillen4 IDM, University of Valencia; 2Department of Burn and Plastic Surgery, La Fe University Hospital; 3Department Orthopaedic Surgery and Traumatology, La Ribera University Hospital; 4IDM, University of Valencia and Department of Pharmacy, CEU-Cardenal Herrera, ValenciaIntroduction: Adipose tissue-derived mesenchymal stem cells (ADMSC) release extracellular vesicles (EV) both below physiological and pathological conditions. The immunomodulatory and anti-inflammatory properties of AD-MSC have confirmed to become effective in several ailments. Osteoarthritis (OA) is usually a major cause of disability in the elderly. Cartilage destruction is mediated by modifications in chondrocyte metabolism and also the up-regulation of inflammatory or catabolic genes. In OA chondrocytes, the induction of autophagy may well be a protective mechanism against strain. We’ve got investigated the effects of microvesicles (MV) and exosomes (EX) from AD-MSC on autophagy, measured as LC3Bpositive autophagosome formation, as well as the production of inflammatory and catabolic mediators in OA chondrocytes stimulated with IL-1. CLL-1 Proteins Recombinant Proteins Techniques: AD-MSC were isolated from fat of individuals who undergone abdominoplasty. EV were isolated from AD-MSC conditioned medium by differential centrifugation with size filtration. Tunable resistive pulse sensing was utilized to evaluate the concentration and size of Ex and Mv. OA chondrocytes had been stimulated with IL-1 (10 ng/mL) and treated with MV (3.6 107 particles/mL) or EX (7.two 107 particles/mL) for 24 h. The levels of oxidised proteins, IL-6, IL-10 and TNF have been measured by ELISA, PGE2 by RIA, and MMP activity and NO by fluorometry. The expression of collagen II and LC3B was evaluated by confocal microscopy. The data had been analysed by ANOVA followed by Dunnett’s test. Benefits and Conclusion: EV down-regulated the production of inflammatory and degradative mediators induced by IL-1. Therapy of OA chondrocytes with MV or EX resulted within a substantial reduction of MMP activity, oxidative strain, IL-6 and TNF levels. In addition, they improved the production of the anti-inflammatory cytokine IL-10 and also the expression of collagen II. Both forms of EV promoted the liberation of LC3B-positive autophagosomes, with a larger effect for MV. Our information indicate that EV exert protective effects on OA chondrocytes and may well have possible pharmacological applications to handle autophagy, inflammatory processes and extracellular matrix degradation. Funding: SAF2013-48724-R (MINECO, FEDER).
