Biomarker Discovery Team, Neurosciences and Signalling Group, Institute of Biomedicine (iBiMED), Department of Medical Sciences, University of Aveiro, 3810-193, Aveiro, Portugal.
Anal Bioanal Chem. 2023 Mar;415(7):1239-1263. doi: 10.1007/s00216-022-04174-5. Epub 2022 Jul 15.
Exosomes emerged as valuable sources of disease biomarkers and new therapeutic tools. However, extracellular vesicles isolation with exosome-like characteristics from certain biofluids is still challenging which can limit their potential use in clinical settings. While ultracentrifugation-based procedures are the gold standard for exosome isolation from cell cultures, no unique and standardized method for exosome isolation from distinct body fluids exists. The complexity, specific composition, and physical properties of each biofluid constitute a technical barrier to obtain reproducible and pure exosome preparations, demanding a detailed characterization of both exosome isolation and characterization methods. Moreover, some isolation procedures can affect downstream proteomic or RNA profiling analysis. This review compiles and discussed a set of comparative studies addressing distinct exosome isolation methods from human biofluids, including cerebrospinal fluid, plasma, serum, saliva, and urine, also focusing on body fluid specific challenges, physical properties, and other potential variation sources. This summarized information will facilitate the choice of exosome isolation methods, based on the type of biological samples available, and hopefully encourage the use of exosomes in translational and clinical research.
外泌体已成为疾病生物标志物和新型治疗工具的有价值来源。然而,从某些生物体液中分离具有外泌体样特征的细胞外囊泡仍然具有挑战性,这可能限制了它们在临床环境中的潜在用途。虽然基于超速离心的程序是从细胞培养物中分离外泌体的金标准,但从不同体液中分离外泌体没有独特和标准化的方法。每种生物体液的复杂性、特定组成和物理性质构成了获得可重复和纯净的外泌体制剂的技术障碍,需要对外泌体分离和表征方法进行详细表征。此外,一些分离程序可能会影响下游蛋白质组学或 RNA 分析。本综述汇集并讨论了一组比较研究,这些研究涉及从人生物体液中分离不同的外泌体方法,包括脑脊液、血浆、血清、唾液和尿液,同时还重点关注体液特有的挑战、物理特性和其他潜在的变化来源。这些汇总信息将有助于根据可用的生物样本类型选择外泌体分离方法,并有望鼓励在外泌体在转化和临床研究中的应用。