Influence of mesenchymal stem cell-derived extracellular vesicles in vitro and their role in ageing

• Published in: Stem cell research & therapy Vol. 11; no. 1; p. 13
• Main Authors: Fafián-Labora, Juan, Morente-López, Miriam, Sánchez-Dopico, María José, Arntz, Onno J, van de Loo, Fons A J, De Toro, Javier, Arufe, María C
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 03.01.2020; BioMed Central; BMC
• Subjects: Ageing; Aging; AKT protein; Analysis; Animals; Annealing; Bone marrow; Diseases; Electron microscopy; Extracellular vesicles; Extracellular Vesicles - metabolism; Mesenchymal stem cell-derived extracellular vesicles; Mesenchymal stem cells; Mesenchymal Stem Cells - metabolism; MicroRNA; MicroRNAs; Microscopy; miRNA; mTOR pathway; Nanoparticles; Oct-4 protein; Penicillin; Pluripotency; Polymerase chain reaction; Rats; Rats, Wistar; Reverse transcription; Scientific equipment industry; Senescence; Signal transduction; Stem cell transplantation; Stem cells; Time; TOR protein; Transmission electron microscopy; Ultracentrifugation; Vinculin; Western blotting; β-Galactosidase; United States; Missouri; Spain; Canada; St Louis Missouri; United Kingdom > UK; United States > US; Japan; France; mTOR pathway; Mesenchymal stem cell-derived extracellular vesicles; Ageing; Pluripotency
• ISSN: 1757-6512; 1757-6512
• DOI: 10.1186/s13287-019-1534-0

This study assessed whether mesenchymal stem cell (MSC)-derived extracellular vesicles influenced ageing and pluripotency markers in cell cultures where they are added. MSC-derived extracellular vesicles from old and young rat bone marrows were isolated by ultracentrifugation and were characterised by western blotting, nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). They were added to young and old MSC cultures. Real-time quantitative reverse transcription polymerase chain reactions and western blot analysis were performed to check the markers of ageing (vinculin and lamin A), pluripotency markers (Nanog and Oct4) and components of the mTOR signalling pathway (Rictor, Raptor, AKT and mTOR) in these cell populations. Subsequently, microRNA (miR)-188-3p expression was transiently inhibited in young MSCs to demonstrate the influence of mTOR2 on MSC ageing. Incubation with young MSC-derived extracellular vesicles decreased the levels of ageing markers and components of the mTOR pathway and increased the pluripotency markers from old MSC populations. By contrast, incubation of young MSCs with old MSC-derived extracellular vesicles generated the reverse effects. Inhibition of miR-188-3p expression in young MSCs produced extracellular vesicles that when incubated with old MSCs produced an increase in the levels of Rictor, as well as a decrease of phosphor-AKT, as indicated by a significant decrease in beta-galactosidase staining. MSC-derived extracellular vesicles affected the behaviour of MSC cultures, based on their composition, which could be modified in vitro. These experiments represented the basis for the development of new therapies against ageing-associated diseases using MSC-derived extracellular vesicles.