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Extracellular vesicles from human plasma for biomarkers discovery: Impact of anticoagulants and isolation techniques.
Bettio, Valentina; Mazzucco, Eleonora; Antona, Annamaria; Cracas, Silvia; Varalda, Marco; Venetucci, Jacopo; Bruno, Stefania; Chiabotto, Giulia; Venegoni, Chiara; Vasile, Alessandra; Chiocchetti, Annalisa; Quaglia, Marco; Camussi, Giovanni; Cantaluppi, Vincenzo; Panella, Massimiliano; Rolla, Roberta; Manfredi, Marcello; Capello, Daniela.
Afiliación
  • Bettio V; Department of Translational Medicine, Center of Excellence in Aging Sciences, University of Piemonte Orientale, Novara, Italy.
  • Mazzucco E; UPO Biobank, University of Piemonte Orientale, Novara, Italy.
  • Antona A; Department of Translational Medicine, Center of Excellence in Aging Sciences, University of Piemonte Orientale, Novara, Italy.
  • Cracas S; UPO Biobank, University of Piemonte Orientale, Novara, Italy.
  • Varalda M; Department of Translational Medicine, Center of Excellence in Aging Sciences, University of Piemonte Orientale, Novara, Italy.
  • Venetucci J; Department of Translational Medicine, Center of Excellence in Aging Sciences, University of Piemonte Orientale, Novara, Italy.
  • Bruno S; Department of Translational Medicine, Center of Excellence in Aging Sciences, University of Piemonte Orientale, Novara, Italy.
  • Chiabotto G; Department of Translational Medicine, Center of Excellence in Aging Sciences, University of Piemonte Orientale, Novara, Italy.
  • Venegoni C; Department of Medical Sciences, University of Torino, Turin, Italy.
  • Vasile A; Department of Medical Sciences, University of Torino, Turin, Italy.
  • Chiocchetti A; Interdisciplinary Research Center of Autoimmune Diseases, Center on Autoimmune and Allergic Diseases, University of Piemonte Orientale, Novara, Italy.
  • Quaglia M; Department of Health Science, "Maggiore della Carità" University Hospital, Novara, Italy.
  • Camussi G; Department of Translational Medicine, Center of Excellence in Aging Sciences, University of Piemonte Orientale, Novara, Italy.
  • Cantaluppi V; Interdisciplinary Research Center of Autoimmune Diseases, Center on Autoimmune and Allergic Diseases, University of Piemonte Orientale, Novara, Italy.
  • Panella M; Department of Health Science, "Maggiore della Carità" University Hospital, Novara, Italy.
  • Rolla R; Department of Translational Medicine, Center of Excellence in Aging Sciences, University of Piemonte Orientale, Novara, Italy.
  • Manfredi M; Nephrology and Kidney Transplantation Unit, Department of Translational Medicine, University of Piemonte Orientale, "Maggiore della Carità" University Hospital, Novara, Italy.
  • Capello D; Department of Medical Sciences, University of Torino, Turin, Italy.
PLoS One ; 18(5): e0285440, 2023.
Article en En | MEDLINE | ID: mdl-37163560
Extracellular vesicles (EVs) isolated from plasma are increasingly recognized as promising circulating biomarkers for disease discovery and progression, as well as for therapeutic drug delivery. The scientific community underlined the necessity of standard operative procedures for the isolation and storage of the EVs to ensure robust results. The understanding of the impact of the pre-analytical variables is still limited and some considerations about plasma anticoagulants and isolation methods are necessary. Therefore, we performed a comparison study between EVs isolated by ultracentrifugation and by affinity substrate separation from plasma EDTA and sodium citrate. The EVs were characterized by Nano Tracking Analysis, Western Blot, cytofluorimetric analysis of surface markers, and lipidomic analysis. While anticoagulants did not significantly alter any of the analyzed parameters, the isolation methods influenced EVs size, purity, surface markers expression and lipidomic profile. Compared to ultracentrifugation, affinity substrate separation yielded bigger particles highly enriched in tetraspanins (CD9, CD63, CD81), fatty acids and glycerolipids, with a predominant LDL- and vLDL-like contamination. Herein, we highlighted that the isolation method should be carefully evaluated prior to study design and the need of standardized operative procedures for EVs isolation and application to biomarkers discovery.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Vesículas Extracelulares / Anticoagulantes Límite: Humans Idioma: En Revista: PLoS One Asunto de la revista: CIENCIA / MEDICINA Año: 2023 Tipo del documento: Article País de afiliación: Italia Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
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XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Vesículas Extracelulares / Anticoagulantes Límite: Humans Idioma: En Revista: PLoS One Asunto de la revista: CIENCIA / MEDICINA Año: 2023 Tipo del documento: Article País de afiliación: Italia Pais de publicación: Estados Unidos