Standardization of sample collection, isolation and analysis methods in extracellular vesicle research

By Kenneth W. Witwer, Edit I. Buzás, L. Bemis, Adriana Bora, Cecilia Lässer, Jan Lötvall, E. N. Nolte-‘t Hoen, Melissa G. Piper, S. Sivaraman, J. Skog and 3 more. Journal of Extracellular Vesicles, 2013

2,322 citations recorded by the source corpus.

The emergence of publications on extracellular RNA (exRNA) and extracellular vesicles (EV) has highlighted the potential of these molecules and vehicles as biomarkers of disease and therapeutic targets. These findings have created a paradigm shift, most prominently in the field of oncology, prompting expanded interest in the field and dedication of funds for EV research. At the same time, understanding of EV subtypes, biogenesis, cargo and mechanisms of shuttling remains incomplete. The techniques that can be harnessed to address the many gaps in our current knowledge were the subject of a special workshop of the International Society for Extracellular Vesicles (ISEV) in New York City in October 2012. As part of the “ISEV Research Seminar: Analysis and Function of RNA in Extracellular Vesicles (evRNA)”, 6 round-table discussions were held to provide an evidence-based framework for isolation and analysis of EV, purification and analysis of associated RNA molecules, and molecular engineering of EV for therapeutic intervention. This article arises from the discussion of EV isolation and analysis at that meeting. The conclusions of the round table are supplemented with a review of published materials and our experience. Controversies and outstanding questions are identified that may inform future research and funding priorities. While we emphasize the need for standardization of specimen handling, appropriate normative controls, and isolation and analysis techniques to facilitate comparison of results, we also recognize that continual development and evaluation of techniques will be necessary as new knowledge is amassed. On many points, consensus has not yet been achieved and must be built through the reporting of well-controlled experiments.

Subfields: Infectious Diseases, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/31bf959a7fc1dde41649ca5d716792b45c33b75b

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Citations in the corpus

Innovative approaches for cancer treatment: current perspectives and new challenges (2019, background): “Specifically, exosomes are small vesicles of around 30–150 nm originated from endosomes in physiological and pathological conditions and released by a fusion of multivesicular bodies (MVBs) to the cell membrane [55, 56], while shed microvesicles (sMVs), with a typical size of 50–1,300 nm, are presen”
Isolation of Extracellular Vesicles: General Methodologies and Latest Trends (2018, background): “Nonetheless, differential centrifugation even when SW rotors are used enables isolation of pure fractions of particles only when they considerably differ in their sedimentation rate, which depends on the buoyant density, that is, the size of EVs and the density of their content [54].”
MIFlowCyt-EV: a framework for standardized reporting of extracellular vesicle flow cytometry experiments (2020, methodology): “General guidelines regarding methods and data reporting have been developed for both flow cytometry (Minimum Information about a Flow Cytometry experiment, MIFlowCyt) [3] and EV research (Minimum Information for the Study of EVs, MISEV) [4,5].”
Manufacturing Exosomes: A Promising Therapeutic Platform. (2018, neutral): “, k-factor or rotor type), meaning that processes are only readily transferable if identical equipment and protocols are used [62,63].”
Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles (2014, neutral): “In the meantime, readers can also refer to 2 detailed Position Papers of ISEV published in 2013, listing recommendations on EV isolation (10) and EV/exRNA analysis (11).”
The impact of disparate isolation methods for extracellular vesicles on downstream RNA profiling (2014, background): “This is a main topic of interest for exosome researchers worldwide, as illustrated by recent position papers by the International Society for Extracellular Vesicles (19,30).”
Exosomes maintain cellular homeostasis by excreting harmful DNA from cells (2017, neutral): “Exosomes are endosomal membrane vesicles with diameters of B 40–150nm 18–20 .”
Biological properties of extracellular vesicles and their physiological functions (2015, background): “Since this early workon blood-derived EVs, it has been recommended by some that EV studies should preferably be conducted on plasma (46), since plasma is the physiological fluid of blood and EVs may potentially be generated in serum after the blood collection during blood clotting (46,315,316).”
Obstacles and opportunities in the functional analysis of extracellular vesicle RNA – an ISEV position paper (2017, background): “[34] Although these are still valid, we have extended”
Overview of Extracellular Vesicles, Their Origin, Composition, Purpose, and Methods for Exosome Isolation and Analysis (2019, background): “These transmembrane proteins, and other proteins associated with the plasma membrane, are commonly found in exosomes and are often enriched in the vesicles compared to the cell lysate [56,57].”