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Platelet removal by single-step centrifugation

机译：单步离心血小板去除

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摘要

The study of extracellular vesicles (EVs) in plasma requires removal of cells including platelets. At present, a two-step centrifugation protocol is recommended and commonly used. A simpler protocol that is less operator dependent is likely to improve the quality of plasma samples collected for EV research. The objective of this study is to develop an easy, fast and clinically applicable centrifugation protocol to produce essentially platelet-free plasma with a high yield for EV research. We compared the two-step centrifugation protocol to a single-step protocol at 5,000 g for 20 minutes. The removal of platelets was computationally predicted and experimentally validated. Flow cytometry was used to detect residual platelets and platelet-derived (CD61+) EVs. The single-step protocol at 5,000 g (i) is less laborious and approximately ten minutes faster, (ii) removes platelets as effective as the two-step centrifugation protocol, and (iii) has a similar to 10% higher plasma yield, whereas (iv) the recovery of platelet-derived EVs is comparable. For future research on plasma EVs we recommend the newly developed, easy and fast single-step protocol for preparation of platelet-free plasma for research on plasma biomarkers including EVs.
机译：研究血浆中的细胞外小泡（EV）需要去除包括血小板在内的细胞。目前，推荐并普遍使用两步离心方案。一个更简单、对操作者依赖性更小的协议可能会改善为EV研究收集的血浆样本的质量。本研究的目的是开发一种简单、快速且临床适用的离心方案，以高产量生产基本上无血小板的血浆，用于EV研究。我们比较了两步离心法和一步离心法，在5000克下离心20分钟。对血小板的去除进行了计算预测和实验验证。流式细胞术用于检测残余血小板和血小板衍生（CD61+）EV。5000 g的单步方案（i）不那么费力，大约快10分钟，（ii）去除血小板的效果与两步离心方案一样有效，（iii）血浆产量高出10%，而（iv）血小板衍生EV的回收率相当。对于血浆EV的未来研究，我们推荐新开发的简单快速的单步无血小板血浆制备方案，用于包括EV在内的血浆生物标记物的研究。

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