In ischemic acute kidney injury Jose Luis Vinas1; Matthew Spence1; Alex Gutsol1; William Knoll1; David Allan2; Burns Kevin1 Kidney Research Centre, Ottawa Hospital Study Institute, University of Ottawa, Ottawa, Canada; 2Ottawa Hospital Investigation Institute, University of Ottawa, Ottawa, CanadaBackground: Infusion of human cord blood endothelial colony forming cell (ECFC)-derived exosomes protects mice against ischaemia/Cystatin B Proteins Source reperfusion acute kidney injury (AKI), by way of transfer of exosomal microRNA(miR)-486-5p. Mechanisms mediating recruitment and retention of exosomes to injured tissues are unclear. The interaction of CXC chemokine receptor form 4 (CXCR4) with stromal cell-derived issue (SDF)-1 has been shown to promoteBackground: Labelling of vesicles for their visualization in vitro or in vivo, involves the usage of fluorescent dyes. To acquire labelled vesicles no cost of unincorporated dye, purification actions are necessary. The regular approach is density gradient ultracentrifugation which is not merely time consuming, but counts with higher sample loss and requires costly equipment. Right here, we established a very simple and speedy approach to acquire labelled vesicles for in vivo tracking and visualization. Methods: Extracellular vesicles (EVs) from cell culture supernatant, synthetic exoliposomes (ELIP) and thermosensitive liposomes (TLIP) were obtained and characterized by nanosight, transmission electron microscopy and zeta possible determinations. Subsequently, the nanostructures have been incubated with DiR fluorophore. DiR-labelled vesicles had been purified by two different methods, using optiprep density gradient ultracentrifugation or commercial exo-spin columns. The eluates obtained from columns and density gradient fractions were characterized by nanosight, dynamic light scattering, zeta prospective, protein content material, fluorescence spectroscopy and imaging. Obtained yields of labelled vesicles were compared. Next, purified labelled EVs, ELIP and TLIP have been administrated by means of tail vein injection in mice with an equivalent number of particles and visualized at 48 h using In Vivo imaging system. Organs have been extracted, visualized and fluorescence intensity was measured. All animal procedures and care had been authorized by implicated ethic committees.Saturday, 05 MayResults: Employing exo-spin column, DiR labelled EVs, ELIP and TLIP had been obtained. Profound characterization of every step, column and ADAMTS Like 5 Proteins Recombinant Proteins eluate through the procedure showed that free DiR was not present in labelled samples. Subsequent, we established that the use of column gives reproducible results with low sample loss. The working time is significantly less than ten min, considerably much less than as much as 24 h in the density gradient system. Finally, we applied these labelled vesicles to determine and evaluate their biodistribution in organs of mice. Summary/Conclusion: We compared two methods and established the use of exo-spin column as a tool to receive labelled vesicles in a reproducible, easy and more quickly manner, with no want of high-priced gear. Funding: This study was funded by FONDECYT 3160592, 11140204, 11150624, 3160323, 1151411, 11140204, and FONDAP 15130011.Centre de recherche d’Organog e Exp imental de l’UniversitLaval/ LOEX, Qu ec, Canada; 2UniversitLaval, Quebec, CanadaPS03.Circulating exosomes as delivery mechanism of cost-free fatty acids (cFFA) Elena Grueso1; Nahuel Aquiles. Garc two; Akaitz Dorronsoro Gonz ez1; Hernan Gonz ez-King1; Rafael S chez1; Alicia Mart ez3; Beatriz J ega4; Enrique O’Connor3; Jose Anastasio Montero1; P.
