Circulating Microparticles: Optimization and Standardization of Isolation Protocols and Reassessment of Their Characteristics and Functions

Microparticles (MPs) are convenient for clinical diagnosis, and have functional roles in signal transduction. Although the importance of MPs is being increasingly recognized, the diversity of isolated protocols for MPs results in a heterogeneous population of their unknown origins, even expands to uncertain functions. Here we systematically studied the composition of MPs at different centrifugal speed intervals, and found that 3000g was a critical centrifugation speed in determining new MPs composition. The platelet-derived particles accounted for more than 80% under 3000g, while only about 20% in MPs obtained over 3000g. Furthermore, we found that the function of new MPs was significantly different from that of traditional ones, such as procoagulation activity, anti-inflammation and clinical diagnosis etc. Thus, our work optimized the method of MPs isolation, clarified some characteristics and physiological functions that should belong to platelets rather than MPs, which will derive new conceptual MPs for its composition and function.

[1]  R. Rakhit,et al.  Persistent circulating platelet and endothelial derived microparticle signature may explain on-going pro-thrombogenicity after acute coronary syndrome. , 2021, Thrombosis research.

[2]  M. Eldh,et al.  Molecular evaluation of five different isolation methods for extracellular vesicles reveals different clinical applicability and subcellular origin , 2021, Journal of extracellular vesicles.

[3]  D. Devine,et al.  Releasates of riboflavin/UV‐treated platelets: Microvesicles suppress cytokine‐mediated endothelial cell migration/proliferation , 2021, Transfusion.

[4]  S. Erzurum,et al.  Characterization and origins of cell-free mitochondria in healthy murine and human blood. , 2020, Mitochondrion.

[5]  G. Mias,et al.  Characterizing Extracellular Vesicles and Their Diverse RNA Contents , 2020, Frontiers in Genetics.

[6]  P. Connes,et al.  Plasma microparticles of sickle patients during crisis or taking hydroxyurea modify endothelium inflammatory properties. , 2020, Blood.

[7]  Gang Wu,et al.  Irradiated tumor cell–derived microparticles mediate tumor eradication via cell killing and immune reprogramming , 2020, Science Advances.

[8]  C. D. Dela Cruz,et al.  A potential role of microvesicle-containing miR-223/142 in lung inflammation , 2019, Thorax.

[9]  G. Raposo,et al.  Extracellular Vesicles: Exosomes and Microvesicles, Integrators of Homeostasis. , 2019, Physiology.

[10]  Changlong Li,et al.  Effect of Endothelial Microparticles Induced by Hypoxia on Migration and Angiogenesis of Human Umbilical Vein Endothelial Cells by Delivering MicroRNA-19b , 2018, Chinese medical journal.

[11]  Xiaoyu Liang,et al.  Circulating Tumor Microparticles Promote Lung Metastasis by Reprogramming Inflammatory and Mechanical Niches via a Macrophage-Dependent Pathway , 2018, Cancer Immunology Research.

[12]  T. Dokland,et al.  Exosomal transfer of mitochondria from airway myeloid-derived regulatory cells to T cells☆ , 2018, Redox biology.

[13]  J. Freedman,et al.  Circulating Platelets as Mediators of Immunity, Inflammation, and Thrombosis , 2018, Circulation research.

[14]  Graça Raposo,et al.  Shedding light on the cell biology of extracellular vesicles , 2018, Nature Reviews Molecular Cell Biology.

[15]  Chang-Qing Zhang,et al.  Platelet-derived Extracellular Vesicles: An Emerging Therapeutic Approach , 2017, International journal of biological sciences.

[16]  L. Edelstein The role of platelet microvesicles in intercellular communication , 2017, Platelets.

[17]  É. Boilard,et al.  Platelet microvesicles in health and disease , 2017, Platelets.

[18]  I. Bruce,et al.  Microparticle subpopulations are potential markers of disease progression and vascular dysfunction across a spectrum of connective tissue disease , 2016, BBA clinical.

[19]  W. Kuebler,et al.  Biomarkers in Lung Diseases : from Pathogenesis to Prediction to New Therapies Microparticles as biomarkers of lung disease : enumeration in biological fluids using lipid bilayer microspheres , 2016 .

[20]  Hong-Jian Zhu,et al.  Extracellular vesicle isolation and characterization: toward clinical application. , 2016, The Journal of clinical investigation.

[21]  Cheuk-Kwan Sun,et al.  Administered circulating microparticles derived from lung cancer patients markedly improved angiogenesis, blood flow and ischemic recovery in rat critical limb ischemia , 2015, Journal of Translational Medicine.

[22]  R. Star,et al.  Microparticles: markers and mediators of sepsis-induced microvascular dysfunction, immunosuppression, and AKI , 2015, Kidney international.

[23]  L. O’Driscoll,et al.  Biological properties of extracellular vesicles and their physiological functions , 2015, Journal of extracellular vesicles.

[24]  Y. Li,et al.  Discovery of the migrasome, an organelle mediating release of cytoplasmic contents during cell migration , 2014, Cell Research.

[25]  D. Devos,et al.  Platelet microparticles: detection and assessment of their paradoxical functional roles in disease and regenerative medicine. , 2014, Blood reviews.

[26]  Romaric Lacroix,et al.  Plasmatic level of leukocyte-derived microparticles is associated with unstable plaque in asymptomatic patients with high-grade carotid stenosis. , 2013, Journal of the American College of Cardiology.

[27]  F. Dignat-George,et al.  Microparticles as a circulating source of procoagulant and fibrinolytic activities in the circulation. , 2012, Thrombosis research.

[28]  R Lacroix,et al.  Impact of pre‐analytical parameters on the measurement of circulating microparticles: towards standardization of protocol , 2012, Journal of thrombosis and haemostasis : JTH.

[29]  Christian Jung,et al.  Circulating endothelial and platelet derived microparticles reflect the size of myocardium at risk in patients with ST-elevation myocardial infarction. , 2012, Atherosclerosis.

[30]  J. Tait,et al.  A new microparticle size calibration standard for use in measuring smaller microparticles using a new flow cytometer , 2011, Journal of thrombosis and haemostasis : JTH.

[31]  H. Tse,et al.  Novel Endothelial Biomarkers: Implications for Periodontal Disease and CVD , 2011, Journal of dental research.

[32]  M. Haubitz,et al.  Detection of circulating microparticles by flow cytometry: influence of centrifugation, filtration of buffer, and freezing , 2010, Vascular health and risk management.

[33]  A. Mügge,et al.  Circulating endothelial microparticles correlate inversely with endothelial function in patients with ischemic left ventricular dysfunction. , 2008, Journal of cardiac failure.

[34]  S. Homer-Vanniasinkam,et al.  Platelet function and antiplatelet therapy , 2007, The British journal of surgery.

[35]  Roeland M. H. Merks,et al.  Endothelial microparticles affect angiogenesis in vitro: role of oxidative stress. , 2005, American journal of physiology. Heart and circulatory physiology.

[36]  Tomohiro Sakamoto,et al.  Elevated levels of VE-cadherin-positive endothelial microparticles in patients with type 2 diabetes mellitus and coronary artery disease. , 2005, Journal of the American College of Cardiology.

[37]  R. Nieuwland,et al.  Measuring circulating cell‐derived microparticles , 2004, Journal of thrombosis and haemostasis : JTH.

[38]  C. Hack,et al.  Human cell‐derived microparticles promote thrombus formation in vivo in a tissue factor‐dependent manner , 2003, Journal of thrombosis and haemostasis : JTH.

[39]  P. Sims,et al.  Complement proteins C5b-9 cause release of membrane vesicles from the platelet surface that are enriched in the membrane receptor for coagulation factor Va and express prothrombinase activity. , 1988, The Journal of biological chemistry.

[40]  P. Wolf The Nature and Significance of Platelet Products in Human Plasma , 1967, British journal of haematology.

[41]  P. Siljander,et al.  Platelet-derived microparticles - an updated perspective. , 2011, Thrombosis research.