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oai:repisalud.isciii.es:20.500.12105/180072024-09-27T12:10:17Zcom_20.500.12105_15322com_20.500.12105_2051col_20.500.12105_16927col_20.500.12105_16967

00925njm 22002777a 4500 dc Riquelme, Jaime A author Takov, Kaloyan author Santiago-Fernández, Concepción author Rossello, Xavier author Lavandero, Sergio author Yellon, Derek M author Davidson, Sean M author 2020-02-26 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. 10.1111/jcmm.15047 1582-4934 Journal of cellular and molecular medicine http://hdl.handle.net/10668/15164 https://hdl.handle.net/20.500.13003/19647 32101370 L2004340775 2-s2.0-85080049735 http://hdl.handle.net/20.500.12105/18007 516109100001 Endothelium Exosomes Extracellular vesicles Senescence Increased production of functional small extracellular vesicles in senescent endothelial cells