Department of Biomolecular Sciences, University of Urbino Carlo Bo, Urbino, Italy.
Methods Mol Biol. 2021;2292:153-172. doi: 10.1007/978-1-0716-1354-2_14.
Extracellular vesicles (EVs) are small membrane-bound particles released into extracellular space by almost all cell types, and found in body fluids like blood, urine, and saliva. Mounting evidence has demonstrated the clinical potential of EVs as diagnostic and therapeutic tools to analyse physiological/pathological processes due to their ability to transport biomolecules secreted from diverse tissues of an individual.For example, the urinary EVs (uEVs), released from all regions of the kidney's nephron and from other cells that line the urinary tract, retain proteomic and transcriptomic markers specific to their cell of origin representing a valuable tool for kidney disease diagnosis.Despite the numerous efforts in developing suitable methods to separate EVs from biofluids, providing material of high purity and low variability poses a limit to clinical translation.This chapter focuses on advantages and disadvantages of several EV isolation methodologies, and provides examples of uEV isolation protocols based on time, cost, and equipment considerations, as well as the sample requirements for any downstream analyses.
细胞外囊泡（EVs）是几乎所有细胞类型释放到细胞外空间的小型膜结合颗粒，存在于血液、尿液和唾液等体液中。越来越多的证据表明，EVs 作为诊断和治疗工具具有临床潜力，可用于分析生理/病理过程，因为它们能够运输个体不同组织分泌的生物分子。例如，从肾脏的肾单位的所有区域以及排列在泌尿道的其他细胞释放的尿细胞外囊泡（uEVs）保留了与其起源细胞特异性的蛋白质组学和转录组学标记物，是一种用于肾脏疾病诊断的有价值的工具。尽管在开发从生物流体中分离 EVs 的合适方法方面做出了许多努力，但提供高纯度和低变异性的材料仍然是临床转化的限制。本章重点介绍了几种 EV 分离方法的优缺点，并提供了基于时间、成本和设备考虑以及任何下游分析的样品要求的 uEV 分离方案的示例。
