Measuring circulating cell‐derived microparticles

Robust Label-free, Quantitative Profiling of Circulating Plasma Microparticle (MP) Associated Proteins

“…We thawed PFP slowly on water/ice, as recommended by Jy and colleagues (39), but later questioned by Trummer et al (40), and very recently confirmed as nondestructive for erythrocyte, platelet and whole plasma derived extracellular vesicles (38). Our quantitative MP proteome assessment corroborated the finding of Lacroix et al that any kind of freezing has an effect on MP integrity (37).…”

“…We used older recommendations for PFP preparation, which were well established in our laboratory and partly supported by a recent study (38), using a high-speed second plasma centrifugation step to prepare PFP from cell-free plasma (39,40). We thawed PFP slowly on water/ice, as recommended by Jy and colleagues (39), but later questioned by Trummer et al (40), and very recently confirmed as nondestructive for erythrocyte, platelet and whole plasma derived extracellular vesicles (38).…”

Centrifugation is a crucial step impacting microparticle measurement

“…During our initial review of the literature, we found a range of centrifugation conditions used to pellet microparticles: 10,000 Â g for 10 minutes [2], 14,000 Â g for 2 minutes [3], through 100,000 Â g for 1 hour [4,5]. Unable to find a consensus, we chose to use 13,000 Â g for 2 minutes to sediment microparticles for two reasons.…”

More on: measuring circulating cell-derived microparticles

“…Hence, in such a multidisciplinary context, the hemostasis laboratory can offer original approaches enabling one to address this issue on quantitative or qualitative bases. This explains the earlier initiative of the In addition to their marker value, MPs can disseminate potent bioactive effectors, including blood-borne tissue factor (TF), the main cellular initiator of the clotting cascade, and procoagulant phosphatidylserine, proinflammatory or apoptogenic mediators, with a central role for the P-selectin pathway in the amplification of the generation of MPs harboring TF [1][2][3][4][5][6][7][8][9]. Hence, a general consensus was reached that MPs are true pathogenic markers of prime interest and, of course, worth standardizing.…”

“…The publication of the Forum ÔMeasuring circulating cellderived microparticlesÕ in the October issue of the Journal of Thrombosis and Haemostasis [1][2][3][4][5][6][7] testifies to the need for the standardization of the determination and characterization of membrane microparticles (MPs). The reviews or original references quoted in the different contributions, to which two other recent relevant syntheses can be added [8,9], highlight the significance of MPs in cardiovascular disorders and in other pathological situations where excessive cell stimulation or death are thought to reflect or account for a substantial part of a deleterious process.…”