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Increased production of functional small extracellular vesicles in senescent endothelial cells

Research output: Contribution to journalArticlepeer-review

Jaime A. Riquelme, Kaloyan Takov, Concepción Santiago-Fernández, Xavier Rossello, Sergio Lavandero, Derek M. Yellon, Sean M. Davidson

Original languageEnglish
Pages (from-to)4871-4876
Number of pages6
Issue number8
Published1 Apr 2020

Bibliographical note

Funding Information: This work was supported by the National Institute for Health Research Biomedical Research Centre (BRC233/CM/SD/101320 to SD) and the British Heart Foundation (PG/18/44/33790 to SD; FS/15/70/32044 to KT), the Comision Nacional de Investigacion Cientifica y Tecnologica (CONICYT, Chile): FONDECYT grant 11181000 to JAR and FONDAP grant 15130011 (to S.L and JAR) and a grant from the ISCIII (Spain) (FI16/00241, MV18/00037) to CSF. The authors would like to thank Mark Turmaine for his technical assistance with electron microscopy experiments. Publisher Copyright: © 2020 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.

King's Authors


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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