Journal ArticleDOI
Comparative proteomics evaluation of plasma exosome isolation techniques and assessment of the stability of exosomes in normal human blood plasma
Hina Kalra,Christopher G. Adda,Michael Liem,Ching-Seng Ang,Adam Mechler,Richard J. Simpson,Mark D. Hulett,Suresh Mathivanan +7 more
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TLDR
The OptiPrepTM density gradient method was superior in isolating pure exosomal populations, devoid of highly abundant plasma proteins, and in the context of cellular uptake, the isolated exosomes were able to fuse with target cells revealing that they were indeed biologically active.Abstract:
Exosomes are nanovesicles released by a variety of cells and are detected in body fluids including blood. Recent studies have highlighted the critical application of exosomes as personalized targeted drug delivery vehicles and as reservoirs of disease biomarkers. While these research applications have created significant interest and can be translated into practice, the stability of exosomes needs to be assessed and exosome isolation protocols from blood plasma need to be optimized. To optimize methods to isolate exosomes from blood plasma, we performed a comparative evaluation of three exosome isolation techniques (differential centrifugation coupled with ultracentrifugation, epithelial cell adhesion molecule immunoaffinity pull-down, and OptiPrep(TM) density gradient separation) using normal human plasma. Based on MS, Western blotting and microscopy results, we found that the OptiPrep(TM) density gradient method was superior in isolating pure exosomal populations, devoid of highly abundant plasma proteins. In addition, we assessed the stability of exosomes in plasma over 90 days under various storage conditions. Western blotting analysis using the exosomal marker, TSG101, revealed that exosomes are stable for 90 days. Interestingly, in the context of cellular uptake, the isolated exosomes were able to fuse with target cells revealing that they were indeed biologically active.read more
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Biological properties of extracellular vesicles and their physiological functions
María Yáñez-Mó,Pia Siljander,Zoraida Andreu,Apolonija Bedina Zavec,Francesc E. Borràs,Edit I. Buzás,Krisztina Buzas,Krisztina Buzas,Enriqueta Casal,Francesco Cappello,Joana Carvalho,Joana Carvalho,Eva Colas,Anabela Cordeiro da Silva,Anabela Cordeiro da Silva,Stefano Fais,Juan M. Falcón-Pérez,Irene M. Ghobrial,Bernd Giebel,Mario Gimona,Mario Gimona,Michael W. Graner,Ihsan Gursel,Mayda Gursel,Niels H. H. Heegaard,Niels H. H. Heegaard,An Hendrix,Peter Kierulf,Katsutoshi Kokubun,Maja Kosanović,Veronika Kralj-Iglič,Eva-Maria Krämer-Albers,Saara Laitinen,Cecilia Lässer,Thomas Lener,Thomas Lener,Erzsébet Ligeti,Aija Linē,Georg Lipps,Alicia Llorente,Jan Lötvall,Mateja Manček-Keber,Antonio Marcilla,María Mittelbrunn,Irina Nazarenko,Esther N. M. Nolte-‘t Hoen,Tuula A. Nyman,Lorraine O'Driscoll,Mireia Olivan,Carla Oliveira,Carla Oliveira,Éva Pállinger,Hernando A. del Portillo,Hernando A. del Portillo,Jaume Reventós,Jaume Reventós,Marina Rigau,Eva Rohde,Eva Rohde,Marei Sammar,Francisco Sánchez-Madrid,Nuno Santarém,Nuno Santarém,Katharina Schallmoser,Katharina Schallmoser,Marie Stampe Ostenfeld,Willem Stoorvogel,Roman Štukelj,Susanne G. van der Grein,M. Helena Vasconcelos,M. Helena Vasconcelos,Marca H. M. Wauben,Olivier De Wever +72 more
TL;DR: A comprehensive overview of the current understanding of the physiological roles of EVs is provided, drawing on the unique EV expertise of academia-based scientists, clinicians and industry based in 27 European countries, the United States and Australia.
Journal ArticleDOI
Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles.
Jan Lötvall,Andrew F. Hill,Fred H. Hochberg,Edit I. Buzás,Dolores Di Vizio,Chris Gardiner,Yong Song Gho,Igor V. Kurochkin,Suresh Mathivanan,Peter J. Quesenberry,Susmita Sahoo,Hidetoshi Tahara,Marca H. M. Wauben,Kenneth W. Witwer,Clotilde Théry +14 more
TL;DR: The International Society for Extracellular Vesicles (ISEV) provides researchers with a minimal set of biochemical, biophysical and functional standards that should be used to attribute any specific biological cargo or functions to EVs.
Journal ArticleDOI
Overview of Extracellular Vesicles, Their Origin, Composition, Purpose, and Methods for Exosome Isolation and Analysis
Laura M. Doyle,Michael Zhuo Wang +1 more
TL;DR: The purpose of this review is to not only introduce the different types of extracellular vesicles but also to summarize their differences and similarities, and discuss different methods of exosome isolation and analysis currently used.
Journal ArticleDOI
Optimized exosome isolation protocol for cell culture supernatant and human plasma
Richard J. Lobb,Melanie Becker,Shu Wen Wen,Christina S. F. Wong,Adrian P. Wiegmans,Antoine Leimgruber,Andreas Möller +6 more
TL;DR: It is shown that concentration of cell culture conditioned media using ultrafiltration devices results in increased vesicle isolation when compared to traditional ultracentrifugation protocols, and it is demonstrated that size exclusion isolation is comparable to density gradient purification of exosomes.
Journal ArticleDOI
Progress in Exosome Isolation Techniques.
TL;DR: An overview of exosome isolation techniques is provided, opening up new perspectives towards the development more innovative strategies and devices for more time saving, cost effective, and efficient isolations ofExosomes from a wide range of biological matrices.
References
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Ceramide Triggers Budding of Exosome Vesicles into Multivesicular Endosomes
Katarina Trajkovic,Chieh Hsu,Salvatore Chiantia,Lawrence Rajendran,Dirk Wenzel,Felix T. Wieland,Petra Schwille,Britta Brügger,Mikael Simons +8 more
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Journal Article
Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers
Journal ArticleDOI
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Mikael Simons,Graça Raposo +1 more
TL;DR: Exosomes transfer not only membrane components but also nucleic acid between different cells, emphasizing their role in intercellular communication.