Increased production of functional small extracellular vesicles in senescent endothelial cells
Small extracellular vesicles (EVs) are novel players in vascular biology. However, a thorough understanding of their production and function remains elusive. Endothelial senescence is a key feature of vascular ageing and thus, is an attractive therapeutic target for the treatment of vascular disease...
| Autores: | , , , , , , |
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| Tipo de recurso: | artículo |
| Fecha de publicación: | 2020 |
| País: | España |
| Institución: | Instituto de Salud Carlos III (ISCIII) |
| Repositorio: | Repisalud |
| Idioma: | inglés |
| OAI Identifier: | oai:repisalud.isciii.es:20.500.12105/18007 |
| Acceso en línea: | http://hdl.handle.net/20.500.12105/18007 |
| Access Level: | acceso abierto |
| Palabra clave: | Endothelium Exosomes Extracellular vesicles Senescence Citometría de Flujo Humanos Biomarcadores Senescencia Celular Vesículas Extracelulares Células Endoteliales de la Vena Umbilical Humana Tetraspanina 29 Tetraspanina 30 beta-Galactosidasa Exosomas Células Endoteliales Proteínas de Unión al GTP rab Biomarkers Cellular Senescence Endothelial Cells Extracellular Vesicles Flow Cytometry Human Umbilical Vein Endothelial Cells Humans Tetraspanin 29 Tetraspanin 30 beta-Galactosidase rab GTP-Binding Proteins rab7 GTP-Binding Proteins |
| Sumario: | Small extracellular vesicles (EVs) are novel players in vascular biology. However, a thorough understanding of their production and function remains elusive. Endothelial senescence is a key feature of vascular ageing and thus, is an attractive therapeutic target for the treatment of vascular disease. In this study, we sought to characterize the EV production of senescent endothelial cells. To achieve this, Human Umbilical Vascular Endothelial Cells (HUVECs) were replicated until they reached senescence, as determined by measurement of Senescence-Associated β-Galactosidase activity via microscopy and flow cytometry. Expression of the endosomal marker Rab7 and the EV marker CD63 was determined by immunofluorescence. Small EVs were isolated by ultracentrifugation and characterized using electron microscopy, nanoparticle tracking analysis and immunoassays to assess morphology, size, concentration and expression of exosome markers CD9 and CD81. Migration of HUVECs in response to EVs was studied using a transwell assay. The results showed that senescent endothelial cells express higher levels of Rab7 and CD63. Moreover, senescent endothelial cells produced higher levels of CD9- and CD81-positive EVs. Additionally, small EVs from both young and senescent endothelial cells promoted HUVEC migration. Overall, senescent endothelial cells produce an increased number of functional small EVs, which may have a role in vascular physiology and disease. |
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