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Journal Article•DOI•

Standardization of pre-analytical variables in plasma microparticle determination: results of the International Society on Thrombosis and Haemostasis SSC Collaborative workshop

Romaric Lacroix1, Coralie Judicone, Micah J. Mooberry2, Mohamed Boucekine, Nigel S. Key2, Françoise Dignat-George1 •
Aix-Marseille University1, University of North Carolina at Chapel Hill2
01 Jun 2013-Journal of Thrombosis and Haemostasis (Wiley-Blackwell)-Vol. 11, Iss: 6, pp 1190-1193
TL;DR: In clinical practice, circulating MP originating from blood and vascular cells are elevated in a variety of prothrombotic and inflammatory disorders, cardiovascular diseases, autoimmune conditions, infectious diseases and cancer.
About: This article is published in Journal of Thrombosis and Haemostasis.The article was published on 2013-06-01 and is currently open access. It has received 315 citations till now. The article focuses on the topics: Cell activation.
Citations
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Journal Article•DOI•
Biogenesis, Secretion, and Intercellular Interactions of Exosomes and Other Extracellular Vesicles

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Marina Colombo1, Graça Raposo, Clotilde Théry•
Curie Institute1
06 Oct 2014-Annual Review of Cell and Developmental Biology
TL;DR: Exosomes were described as vesicles of endosomal origin secreted from reticulocytes in the 1980s as discussed by the authors, and their biogenesis, their secretion, and their subsequent fate are discussed, as their functions rely on these important processes.
Abstract: In the 1980s, exosomes were described as vesicles of endosomal origin secreted from reticulocytes. Interest increased around these extracellular vesicles, as they appeared to participate in several cellular processes. Exosomes bear proteins, lipids, and RNAs, mediating intercellular communication between different cell types in the body, and thus affecting normal and pathological conditions. Only recently, scientists acknowledged the difficulty of separating exosomes from other types of extracellular vesicles, which precludes a clear attribution of a particular function to the different types of secreted vesicles. To shed light into this complex but expanding field of science, this review focuses on the definition of exosomes and other secreted extracellular vesicles. Their biogenesis, their secretion, and their subsequent fate are discussed, as their functions rely on these important processes.

3,959 citations

Biogenesis, Secretion, and Intercellular Interactions of Exosomes and Other

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Extracellular Vesicles, Graça Raposo
01 Jan 2014
TL;DR: The definition of exosomes and other secreted extracellular vesicles, which mediating intercellular communication between different cell types in the body, and thus affecting normal and pathological conditions are focused on.
Abstract: In the 1980s, exosomes were described as vesicles of endosomal origin secreted from reticulocytes. Interest increased around these extracellular vesicles, as they appeared to participate in several cellular processes. Exosomes bear proteins, lipids, and RNAs, mediating intercellular communication between different cell types in the body, and thus affecting normal and pathological conditions. Only recently, scientists acknowledged the difficulty of separating exosomes from other types of extracellular vesicles, which precludes a clear attribution of a particular function to the different types of secreted vesicles. To shed light into this complex but expanding field of science, this review focuses on the definition of exosomes and other secreted extracellular vesicles. Their biogenesis, their secretion, and their subsequent fate are discussed, as their functions rely on these important processes.

3,321 citations

Cites background from "Standardization of pre-analytical v..."

  • ...…surface markers on circulating microparticles (Nieuwland et al. 1997); however, most routinely used flow cytometers do not properly distinguish between noise and beads (or vesicles) of sizes below 300 nm and do not separate beads with size differences lower than 200 nm (Lacroix et al. 2013)....

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Journal Article•DOI•
Standardization of sample collection, isolation and analysis methods in extracellular vesicle research

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Kenneth W. Witwer1, Edit I. Buzás2, Lynne T. Bemis3, Adriana Bora4, Cecilia Lässer5, Jan Lötvall5, Esther Nolte-‘t Hoen6, Melissa G. Piper7, Sarada Sivaraman8, Johan Skog, Clotilde Théry9, Marca H. M. Wauben6, Fred H. Hochberg8 •
Johns Hopkins University School of Medicine1, Semmelweis University2, University of Minnesota3, Johns Hopkins University4, University of Gothenburg5, Utrecht University6, Ohio State University7, Harvard University8, Curie Institute9
27 May 2013-Journal of extracellular vesicles
TL;DR: The need for standardization of specimen handling, appropriate normative controls, and isolation and analysis techniques to facilitate comparison of results is emphasized, and it is recognized that continual development and evaluation of techniques will be necessary as new knowledge is amassed.
Abstract: 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. Keywords: extracellular vesicle; exosome; microvesicle; standardization; isolation (Published: 27 May 2013) Citation: Journal of Extracellular Vesicles 2013, 2 : 20360 - http://dx.doi.org/10.3402/jev.v2i0.20360

1,840 citations

Journal Article•DOI•
Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

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Andrea Cossarizza1, Hyun-Dong Chang, Andreas Radbruch, Andreas Acs2  +459 more•Institutions (160)
01 Oct 2019-European Journal of Immunology
TL;DR: These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community providing the theory and key practical aspects offlow cytometry enabling immunologists to avoid the common errors that often undermine immunological data.
Abstract: These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion.

698 citations

Journal Article•DOI•
Methodological Guidelines to Study Extracellular Vesicles.

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Frank A. W. Coumans1, Alain Brisson2, Edit I. Buzás3, Françoise Dignat-George4, Esther E.E. Drees, Samir El-Andaloussi5, Samir El-Andaloussi6, Costanza Emanueli7, Costanza Emanueli8, Aleksandra Gasecka9, Aleksandra Gasecka1, An Hendrix10, Andrew F. Hill11, Romaric Lacroix4, Yi Lee5, Ton G. van Leeuwen1, Nigel Mackman12, Imre Mäger6, Imre Mäger13, John P. Nolan, Edwin van der Pol1, D. Michiel Pegtel, Susmita Sahoo14, Pia Siljander15, Guus Sturk1, Olivier De Wever10, Rienk Nieuwland1 •
University of Amsterdam1, University of Bordeaux2, Semmelweis University3, French Institute of Health and Medical Research4, Karolinska Institutet5, University of Oxford6, University of Bristol7, Imperial College London8, Medical University of Warsaw9, Ghent University10, La Trobe University11, University of North Carolina at Chapel Hill12, University of Tartu13, Icahn School of Medicine at Mount Sinai14, University of Helsinki15
12 May 2017-Circulation Research
TL;DR: This review provides an expert-based update of recent advances in the methods to study EVs and summarizes currently accepted considerations and recommendations from sample collection to isolation, detection, and characterization of EVs.
Abstract: Owing to the relationship between extracellular vesicles (EVs) and physiological and pathological conditions, the interest in EVs is exponentially growing. EVs hold high hopes for novel diagnostic and translational discoveries. This review provides an expert-based update of recent advances in the methods to study EVs and summarizes currently accepted considerations and recommendations from sample collection to isolation, detection, and characterization of EVs. Common misconceptions and methodological pitfalls are highlighted. Although EVs are found in all body fluids, in this review, we will focus on EVs from human blood, not only our most complex but also the most interesting body fluid for cardiovascular research.

651 citations

Cites methods from "Standardization of pre-analytical v..."

  • ...Several anticoagulants have been used to collect blood for analysis of EVs, including EDTA, sodium fluoride/potassium oxalate (NaF/KOx), or (trisodium) citrate.28,36,37 At present, citrate (0.109 mol/L final concentration) is the most commonly used anticoagulant and has been recommended by the International Society on Thrombosis and Haemostasis.25 Both acid citrate dextrose and citrate, theophylline, adenosine and dipyridamole prevent platelet activation and the release of platelet EVs more efficiently than citrate.26,38,39 The choice of anticoagulant strongly depends on the downstream analysis, and, for example, EDTA is a suitable anticoagulant for RNA analysis,40,41 whereas heparin interferes with polymerase chain reaction (PCR).42 Taken together, both the extent of inhibition of EV release in collected blood samples ex vivo and the intended downstream assays should be taken into account when choosing an anticoagulant....

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  • ...anticoagulant and has been recommended by the International Society on Thrombosis and Haemostasis.(25) Both acid citrate...

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References
More filters
Journal Article•DOI•
Circulating microparticles: pathophysiology and clinical implications

[...]

Andrea Piccin, William G. Murphy, Owen P. Smith
01 May 2007-Blood Reviews
TL;DR: The genesis and role of microparticles, derived from platelets, endothelial cells and monocytes, in sepsis, heparin-induced thrombocytopenia, TTP, aplastic anaemia, paroxysmal nocturnal haemoglobinuria, and sickle cell disease have been well studied, and provide important insights into the underlying diseases.

736 citations

"Standardization of pre-analytical v..." refers background in this paper

  • ...In clinical practice, circulating MP originating from blood and vascular cells are elevated in a variety of prothrombotic and inflammatory disorders, cardiovascular diseases, autoimmune conditions, infectious diseases and cancer [1-3]...

    [...]

  • ...1–1 μm membrane particles that expose the anionic phospholipid phosphatidylserine (PS) and membrane antigens representative of their cellular origin [1]....

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  • ...1 to 1 μm membrane particles that expose the anionic phospholipid phosphatidylserine (PS) and membrane antigens representative of their cellular origin [1]....

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Journal Article•DOI•
Measuring circulating cell-derived microparticles

[...]

W. Jy1, Lawrence L. Horstman1, Joaquin J. Jimenez1, Yeon S. Ahn1, E. Biro2, Rienk Nieuwland2, Auguste Sturk2, Françoise Dignat-George3, Florence Sabatier3, Laurence Camoin-Jau3, José Sampol3, B. Hugel4, Fatiha Zobairi4, Jean Marie Freyssinet4, Shosaku Nomura5, Arun S. Shet6, Arun S. Shet7, Nigel S. Key7, Robert P. Hebbel •
University of Miami1, University of Amsterdam2, French Institute of Health and Medical Research3, University of Strasbourg4, Kansai Medical University5, Rockefeller University6, University of Minnesota7
01 Oct 2004-Journal of Thrombosis and Haemostasis
TL;DR: Major differences exist in the preparation of the MP samples, whether or not they are first sedimented and resuspended, means of generic MP detection, and cell lineage-specific antigenic markers, and these differences probably account for some of the different findings among the groups.

442 citations

"Standardization of pre-analytical v..." refers methods in this paper

  • ...Therefore, the main objective of this new workshop was to determine whether it is possible to reduce the inter-laboratory variability in MP count and procoagulant activity using a common pre-analytical protocol [9] compared with a non-standardized approach....

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Journal Article•DOI•
Standardization of platelet-derived microparticle enumeration by flow cytometry with calibrated beads: results of the International Society on Thrombosis and Haemostasis SSC Collaborative workshop.

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Romaric Lacroix1, Stéphane Robert1, Philippe Poncelet, Raj S. Kasthuri2, Nigel S. Key2, Françoise Dignat-George1 •
Aix-Marseille University1, University of North Carolina at Chapel Hill2
01 Nov 2010-Journal of Thrombosis and Haemostasis
TL;DR: A project aimed at standardizing the enumeration of cellularMPs by FCM by checking whether the instrument to be used to enumerate PMPs demonstrated the required performance with a blend of fluorescent beads with well-known sizes and relative amounts and defining the interinstrument reproducibility of PMP enumeration in human plasma.

339 citations

"Standardization of pre-analytical v..." refers background in this paper

  • ...Accordingly, the International Society on Thrombosis and Haemostasis (ISTH) Vascular Biology Standardisation Subcommittee (VB SSC) previously organized a first collaborative working party aimed at standardizing platelet-derived MP (PMP) measurement by flow cytometry [4]....

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Journal Article•DOI•
Impact of pre-analytical parameters on the measurement of circulating microparticles: towards standardization of protocol.

[...]

Romaric Lacroix1, Coralie Judicone, Philippe Poncelet, Stéphane Robert1, Laurent Arnaud, José Sampol1, Françoise Dignat-George1 •
Aix-Marseille University1
01 Mar 2012-Journal of Thrombosis and Haemostasis
TL;DR: Microparticles received an increasing interest both as biomarkers and biovectors in coagulation, inflammation and cancer, and among the limitation of such studies, pre‐analytical steps remains an important source of variability and artifacts in MP analysis.

315 citations

"Standardization of pre-analytical v..." refers background in this paper

  • ...Other variables have been shown to have a significant impact on MP pre-analytics, such as the time before the first centrifugation and the agitation of the tube transportation [8]; however, both these variables have been well controlled during this study....

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Journal Article•DOI•
Pre-analytical and analytical issues in the analysis of blood microparticles

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Yuana Yuana1, Rogier M. Bertina, Susanne Osanto2•
Leiden University1, Leiden University Medical Center2
21 Dec 2010-Thrombosis and Haemostasis
TL;DR: Standardisation of pre-analytical procedures and the introduction of reliable and rapid methods for the measurement of MPs are urgently needed to facilitate their use as biomarker in the pathophysiology of diseases.
Abstract: Results of plasma microparticles (MPs) measurements reported in the literature vary widely. This is clearly not only related to the lack of well-standardised MP assays, but also to variations in pre-analytical conditions. In this review we will discuss the pre-analytical variables related to plasma and MP preparation which may affect MP analysis. Additionally we will address several analytical issues in commonly used MP assays and briefly discuss some novel approaches for the detection and characterisation of MPs. Ideally MP measurements should be performed in plasma, freshly prepared directly after blood withdrawal. As platelet contamination seems to be one of the major pre-analytical problems in processing plasma for MP measurement, the use of platelet-free plasma may be preferred. When frozen-thawed plasma is used, especially PMP and annexinV-positive MP counts should be interpreted with caution. When flow cytometry is chosen as a method for quantification of MPs, some analytical conditions should be standardised, e.g. settings of the flow cytometer, quality of the antibodies, and use of counting beads. Fluorescence-nanoparticle tracking analysis and atomic force microscopy can accurately count nanosized MPs, but unfortunately the operational procedures of both methods are still time consuming and they give no information on the functional properties of MPs. The MP-TF activity assay provides information on MPs carrying active TF, regardless of their parental origin. Ultimately, standardisation of pre-analytical procedures and the introduction of reliable and rapid methods for the measurement of MPs are urgently needed to facilitate their use as biomarker in the pathophysiology of diseases.

268 citations

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