Large‐scale production of extracellular vesicles: Report on the “massivEVs” ISEV workshop (original) (raw)
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International Society for Extracellular Vesicles: first annual meeting, April 17-21, 2012: ISEV-2012
2012
Extracellular micro-and nano-scale membrane vesicles produced by different cells are recognised as an essential entity of physiological fluids in a variety of organisms and function as mediators of intercellular communication employed for the regulation of multiple systemic and local processes. In the last decade, an exponential amount of experimental work was dedicated to exploring the biogenesis and secretion mechanisms, physiological and pathological functions and potential applications of the extracellular vesicles (EVs). Noteworthy is the large heterogeneity of in vitro and in vivo models applied, technical approaches developed in these studies and the diversity of designations assigned to different or similar types of EVs. Hence, there is a clear necessity for a uniform nomenclature and standardisation of methods to isolate and characterise these vesicles. In April 2012, the first meeting of the International Society for Extracellular Vesicles (ISEV) took place bringing together this exponentially grown scientific community. The
2018
The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, andmany other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles (“MISEV”) guidelines for the field in 2014. We now update these “MISEV2014” guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities. Finally, a checklist is provided with summaries of key points.
Biotechnology Journal
Potential applications of extracellular vesicles (EVs) are attracting increasing interest in the fields of medicine, cosmetics, and nutrition. However, the manufacturing of EVs is currently characterized by low yields. This limitation severely hampers progress in research at the laboratory and clinical scales, as well as the realization of successful and cost-effective EV-based products. Moreover, the high level of heterogeneity of EVs further complicates reproducible manufacturing on a large scale. In this review, possible directions toward the scalable production of EVs are discussed. In particular, two strategies are considered: i) the optimization of upstream unit operations and ii) the exploitation of well-established and mature technologies already in use in other industrial bioprocesses.
Taking in the trash: bioreactors for the mass production of clinical-grade extracellular vesicles
The Journal of Cardiovascular Aging, 2022
In recent years, the quest for the next generation of therapeutics for cardiovascular diseases has focused heavily on extracellular vesicles (EVs), and for good reason. Formed by the inward budding of the endosomal membrane, EVs are small secreted vesicles about 50-150 nm in diameter, containing a myriad of bioactive cargo, including lipids, proteins, and micro-RNAs (miRs). Though their existence has been known since at least the early 1980s, EVs were originally considered little more than “garbage bins”, or a means by which cells package and expel unnecessary proteins and other molecules. In the intervening years, an increasing body of evidence has identified EVs as a primary mediator of paracrine signaling between cells. Secreted by nearly every type of cell in the body, EVs have been implicated both in normal physiological processes, such as inflammation and cell proliferation, as well as the pathogenesis of various disease states.
Rigor and standardization of extracellular vesicle research: Paving the road towards robustness
Journal of Extracellular Vesicles, 2020
Accompanying the growing awareness that extracellular vesicles (EVs) are useful for diagnostics and therapeutics is the realization that EV applications must be established with rigor, reproducibility, and standardization. The small size and heterogeneity of most EVs are well-known barriers to rigorous studies. However, progress is also hampered by the largely unknown influence of a host of pre-analytical variables and a lack of quality controls, differences in EV separation and characterization techniques (van Deun et al., 2017), a general lack of dedicated reference materials and standards (Welsh et al., 2020), and analyses that are poorly defined and thus irreproducible. The goal of this Editorial is to summarize the past, present, and future contributions of the International Society for Extracellular Vesicles (ISEV) to improving rigor and standardization in EV research. We highlight in particular the Rigor and Standardization Subcommittee and the recently published results of a survey on the methods used for separation and characterization of EVs (Royo, Théry, Falcón-Pérez, Nieuwland, & Witwer, 2020). Major challenges remain. Even so, we predict that within a few years, the expansion of an ISEV infrastructure dedicated to rigor and standardization will enable us to turn the ugly duckling of current challenges into a beautiful swan of comparable and reproducible EV measurement results. Because we are working in a new and developing field of research, ISEV has the unique opportunity to set high standards for rigor, and we may even lead the way for other research fields and societies. Rigor and standardization with community input have been part of the ISEV mission since its founding. Impetus for founding a society arose with a 2011 meeting in Paris that was organized by Clotilde Théry and Graça Raposo. The incipient community voted to emphasize ' extracellular vesicles' in the name of the society rather than limiting focus to specific subtypes of EVs. Following the first annual meeting of ISEV in 2012, the first ISEV Workshop was organized and held in October, 2012 in New York City. This Workshop gave rise to the first two ISEV position papers, on standardization of sample collection and processing (Witwer et al., 2013) and RNA analysis (Hill et al., 2013). Subsequent workshops and co-sponsored events have given birth to additional position papers and other products, most of them published in the
2018
The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles (“MISEV”) guidelines for the field in 2014. We now update these “MISEV2014” guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond me...
Production of Extracellular Vesicles Loaded with Therapeutic Cargo
Methods in molecular biology (Clifton, N.J.), 2018
Extracellular vesicles (EVs) are biological nanoparticles comprising exosomes, microvesicles, and other heterogeneous nanoscopic vesicle populations that are produced by most cell types. In addition to their putative roles as critical mediators of intercellular communication, EVs have begun to be harnessed as drug delivery vehicles, with early evidence indicating they may have significant advantages over synthetic nanoparticle delivery systems for particular applications. Targeted delivery of EV-encapsulated cargo has already been realized and may have broad applicability; however, methods for producing and purifying EVs and loading them with therapeutic molecules have yet to be standardized. In this chapter, we outline steps for EV isolation and characterization and compare current methods for active and passive loading of EVs with payloads of short interfering RNA (siRNA) or small molecules, with the results revealing that active loading via electroporation increases loading effic...
A brief history of nearly EV‐erything – The rise and rise of extracellular vesicles
Journal of extracellular vesicles, 2021
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