Influence of mesenchymal stem cell-derived extracellular vesicles in vitro and their role in ageing
Influence of mesenchymal stem cell-derived extracellular vesicles in vitro and their role in ageing
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DOI：
10.1186/s13287-019-1534-0
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发表时间：
2020-01-03
影响因子：
7.5
通讯作者：
Arufe, Maria C.
中科院分区：
文献类型：
--
作者：
Fafian-Labora, Juan;Morente-Lopez, Miriam;Arufe, Maria C.
IntroductionThis study assessed whether mesenchymal stem cell（MSC）-derived extracellular vesicles influenced aging and pluripotency marks in cell cultures where they are address. MethodsMSC-derived extracellular vesicles from old and young rat bone marrows were isolated by ultracentbridging and were characterised by western blotting，nanoparticle tracking analysis（NTA）and transmission electron microscopy（TEM）.它们被添加到年轻和年老的MSC培养物中。进行实时定量逆转录聚合酶链反应和蛋白质印迹分析，以检查这些细胞群中的衰老标志物（黏着斑蛋白和核纤层蛋白A）、多能性标志物（Nanog和Oct 4）和mTOR信号通路的组分（Rictor、Raptor、AKT和mTOR）。随后，microRNA（miR）-188-3p的表达被短暂抑制在年轻的MSC表现出mTOR 2对MSC aging.ResultsIncubation的影响与年轻的MSC衍生的细胞外囊泡的老化标志物和组件的mTOR通路的水平降低，并增加了多能性标记从老MSC种群。相比之下，年轻的MSC与旧的MSC衍生的细胞外囊泡孵育产生相反的效果。在年轻的骨髓间充质干细胞中的miR-188- 3 p表达的抑制产生的细胞外囊泡，当与老骨髓间充质干细胞孵育产生的Rictor的水平增加，以及磷酸化AKT的减少，如在β-半乳糖苷酶staining. ConclusionsMSC-衍生的细胞外囊泡的影响MSC培养物的行为，根据其组成，可以在体外进行修改。这些实验代表了使用MSC衍生的细胞外囊泡开发抗衰老相关疾病的新疗法的基础。
IntroductionThis study assessed whether mesenchymal stem cell (MSC)-derived extracellular vesicles influenced ageing and pluripotency markers in cell cultures where they are added.MethodsMSC-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.ResultsIncubation 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.ConclusionsMSC-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.