Low-density lipoprotein mimics blood plasma-derived exosomes and microvesicles during isolation and detection.
Low-density lipoprotein mimics blood plasma-derived exosomes and microvesicles during isolation and detection.
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DOI：
10.1038/srep24316
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发表时间：
2016-04-18
影响因子：
4.6
通讯作者：
Buzás EI
中科院分区：
文献类型：
--
作者：
Sódar BW;Kittel Á;Pálóczi K;Vukman KV;Osteikoetxea X;Szabó-Taylor K;Németh A;Sperlágh B;Baranyai T;Giricz Z;Wiener Z;Turiák L;Drahos L;Pállinger É;Vékey K;Ferdinandy P;Falus A;Buzás EI
循环细胞外囊泡已成为各种疾病中潜在的新生物标志物。尽管兴趣越来越大，但从体液中分离和纯化它们仍然具有挑战性。在这里，我们研究了人餐前和餐后4小时无血小板血浆样本以及人血小板浓缩物。使用流式细胞术，我们发现，大多数循环颗粒的大小范围内的细胞外囊泡缺乏共同的囊泡标记。我们确定这些颗粒中的大多数为脂蛋白（主要是低密度脂蛋白，LDL），其模仿细胞外囊泡的特征，并与它们共纯化。基于LDL的生物物理特性，这一发现是非常出乎意料的。目前最先进的细胞外囊泡分离和纯化方法不能从血浆或血小板浓缩物中获得无脂蛋白的囊泡制剂。此外，透射电子显微镜显示协会的LDL与孤立的囊泡后，在体外混合。这是第一项研究，以显示共纯化和在体外协会的低密度脂蛋白与细胞外囊泡和其干扰囊泡分析。我们的数据表明，在使用血液来源的细胞外囊泡的研究中，特别关注潜在的共纯化LDL，仔细的研究设计和数据解释非常重要。
Circulating extracellular vesicles have emerged as potential new biomarkers in a wide variety of diseases. Despite the increasing interest, their isolation and purification from body fluids remains challenging. Here we studied human pre-prandial and 4 hours postprandial platelet-free blood plasma samples as well as human platelet concentrates. Using flow cytometry, we found that the majority of circulating particles within the size range of extracellular vesicles lacked common vesicular markers. We identified most of these particles as lipoproteins (predominantly low-density lipoprotein, LDL) which mimicked the characteristics of extracellular vesicles and also co-purified with them. Based on biophysical properties of LDL this finding was highly unexpected. Current state-of-the-art extracellular vesicle isolation and purification methods did not result in lipoprotein-free vesicle preparations from blood plasma or from platelet concentrates. Furthermore, transmission electron microscopy showed an association of LDL with isolated vesicles upon in vitro mixing. This is the first study to show co-purification and in vitro association of LDL with extracellular vesicles and its interference with vesicle analysis. Our data point to the importance of careful study design and data interpretation in studies using blood-derived extracellular vesicles with special focus on potentially co-purified LDL.