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Characterization of Plasma-Derived Extracellular Vesicles Isolated by Different Methods: A Comparison Study

机译:不同方法分离的血浆来源细胞外囊的表征:对比研究

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摘要

Extracellular vesicles (EV) are small membrane structures released by cells that act as potent mediators of intercellular communication. The study of EV biology is important, not only to strengthen our knowledge of their physiological roles, but also to better understand their involvement in several diseases. In the field of biomedicine they have been studied as a novel source of biomarkers and drug delivery vehicles. The most commonly used method for EV enrichment in crude pellet involves serial centrifugation and ultracentrifugation. Recently, different protocols and techniques have been developed to isolate EV that imply less time and greater purification. Here we carry out a comparative analysis of three methods to enrich EV from plasma of healthy controls: ultracentrifugation, ExoQuickTM precipitation solution (System Biosciences), and Total Exosome Isolation kit (Invitrogen). Our results show that commercial precipitation reagents are more efficient and enable higher EV enrichment factors compared with traditional ultracentrifugation, although subsequent imaging analysis is not possible with some of them. We hope that this work will contribute to the current research on isolation techniques to assist the progress of clinical applications with diagnostic or therapeutic objectives.
机译:细胞外囊泡(EV)是由细胞释放的小膜结构,可充当细胞间通讯的有效介体。电动汽车生物学的研究很重要,不仅是要增强我们对其生理作用的认识,而且要更好地了解其参与多种疾病。在生物医学领域,它们已被研究为生物标志物和药物递送载体的新来源。粗颗粒中EV富集最常用的方法包括连续离心和超速离心。近来,已经开发出不同的方案和技术来分离EV,这意味着更少的时间和更大的纯化。在这里,我们对三种从健康对照血浆中富集EV的方法进行了比较分析:超速离心,ExoQuick TM 沉淀溶液(System Biosciences)和总外泌体分离试剂盒(Invitrogen)。我们的结果表明,与传统的超速离心相比,商业沉淀试剂更有效,并且能够实现更高的EV富集因子,尽管其中某些试剂无法进行后续的成像分析。我们希望这项工作将有助于当前有关隔离技术的研究,以帮助实现具有诊断或治疗目的的临床应用。

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