• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

一种从血浆或细胞培养基中分离细胞外囊泡的新颖三步方案，可实现高产量和高纯度。

A novel three step protocol to isolate extracellular vesicles from plasma or cell culture medium with both high yield and purity.

作者信息

Zhang Xiaogang, Borg Ellen G F, Liaci A Manuel, Vos Harmjan R, Stoorvogel Willem

机构信息

Department of Biomolecular Health Sciences, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.

Bijvoet Center for Biomolecular Research, Department of Chemistry, Utrecht University, Utrecht, The Netherlands.

出版信息

J Extracell Vesicles. 2020 Jul 14;9(1):1791450. doi: 10.1080/20013078.2020.1791450.

DOI:10.1080/20013078.2020.1791450
PMID:32944179
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7480457/
Abstract

Extracellular vesicles (EV) are membrane encapsulated nanoparticles that can function in intercellular communication, and their presence in biofluids can be indicative for (patho)physiological conditions. Studies aiming to resolve functionalities of EV or to discover EV-associated biomarkers for disease in liquid biopsies are hampered by limitations of current protocols to isolate EV from biofluids or cell culture medium. EV isolation is complicated by the >10-fold numerical excess of other types of particles, including lipoproteins and protein complexes. In addition to persisting contaminants, currently available EV isolation methods may suffer from inefficient EV recovery, bias for EV subtypes, interference with the integrity of EV membranes, and loss of EV functionality. In this study, we established a novel three-step non-selective method to isolate EV from blood or cell culture media with both high yield and purity, resulting in 71% recovery and near to complete elimination of unrelated (lipo)proteins. This EV isolation procedure is independent of ill-defined commercial kits, and apart from an ultracentrifuge, does not require specialised expensive equipment.

摘要

细胞外囊泡（EV）是膜包裹的纳米颗粒，可在细胞间通讯中发挥作用，它们在生物流体中的存在可指示（病理）生理状况。旨在解析EV功能或在液体活检中发现与疾病相关的EV生物标志物的研究受到当前从生物流体或细胞培养基中分离EV的方案的限制。EV的分离因其他类型颗粒（包括脂蛋白和蛋白质复合物）在数量上超过其10倍以上而变得复杂。除了持续存在的污染物外，目前可用的EV分离方法可能存在EV回收率低、对EV亚型有偏差、干扰EV膜的完整性以及EV功能丧失等问题。在本研究中，我们建立了一种新颖的三步非选择性方法，可从血液或细胞培养基中高效、高纯度地分离EV，回收率达71%，几乎完全消除了无关的（脂）蛋白。这种EV分离程序不依赖于不明确的商业试剂盒，除了超速离心机外，不需要专门的昂贵设备。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f7f/7480457/e28afba93936/ZJEV_A_1791450_F0006_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f7f/7480457/c1dbb5a5bfe0/ZJEV_A_1791450_F0001_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f7f/7480457/ebcb6af54bb6/ZJEV_A_1791450_F0002_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f7f/7480457/14c3ec612b2b/ZJEV_A_1791450_F0003_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f7f/7480457/8468e8502519/ZJEV_A_1791450_F0004_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f7f/7480457/1974b276c9cf/ZJEV_A_1791450_F0005_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f7f/7480457/e28afba93936/ZJEV_A_1791450_F0006_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f7f/7480457/c1dbb5a5bfe0/ZJEV_A_1791450_F0001_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f7f/7480457/ebcb6af54bb6/ZJEV_A_1791450_F0002_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f7f/7480457/14c3ec612b2b/ZJEV_A_1791450_F0003_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f7f/7480457/8468e8502519/ZJEV_A_1791450_F0004_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f7f/7480457/1974b276c9cf/ZJEV_A_1791450_F0005_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f7f/7480457/e28afba93936/ZJEV_A_1791450_F0006_OC.jpg

相似文献

1
A novel three step protocol to isolate extracellular vesicles from plasma or cell culture medium with both high yield and purity.一种从血浆或细胞培养基中分离细胞外囊泡的新颖三步方案，可实现高产量和高纯度。
J Extracell Vesicles. 2020 Jul 14;9(1):1791450. doi: 10.1080/20013078.2020.1791450.
2
Comparison of membrane affinity-based method with size-exclusion chromatography for isolation of exosome-like vesicles from human plasma.基于膜亲和力的方法与排阻色谱法比较，用于从人血浆中分离类外泌体囊泡。
J Transl Med. 2018 Jan 9;16(1):1. doi: 10.1186/s12967-017-1374-6.
3
A magnetic bead-mediated selective adsorption strategy for extracellular vesicle separation and purification.一种用于细胞外囊泡分离和纯化的磁珠介导的选择性吸附策略。
Acta Biomater. 2021 Apr 1;124:336-347. doi: 10.1016/j.actbio.2021.02.004. Epub 2021 Feb 10.
4
An Isolation System to Collect High Quality and Purity Extracellular Vesicles from Serum.一种从血清中收集高质量和高纯度细胞外囊泡的分离系统。
Int J Nanomedicine. 2021 Sep 29;16:6681-6692. doi: 10.2147/IJN.S328325. eCollection 2021.
5
Isolation of extracellular vesicles from human plasma samples: The importance of controls.从人血浆样本中分离细胞外囊泡：对照的重要性。
Biotechnol J. 2023 Jun;18(6):e2200575. doi: 10.1002/biot.202200575. Epub 2023 Apr 9.
6
Isolation of High-Purity Extracellular Vesicles by the Combination of Iodixanol Density Gradient Ultracentrifugation and Bind-Elute Chromatography From Blood Plasma.通过碘克沙醇密度梯度超速离心和结合洗脱色谱法从血浆中分离高纯度细胞外囊泡。
Front Physiol. 2018 Oct 23;9:1479. doi: 10.3389/fphys.2018.01479. eCollection 2018.
7
High throughput and rapid isolation of extracellular vesicles and exosomes with purity using size exclusion liquid chromatography.使用尺寸排阻液相色谱法高通量且快速地分离出具有纯度的细胞外囊泡和外泌体。
Bioact Mater. 2024 Sep 4;40:683-695. doi: 10.1016/j.bioactmat.2024.08.002. eCollection 2024 Oct.
8
The polysaccharide chitosan facilitates the isolation of small extracellular vesicles from multiple biofluids.壳聚糖有助于从小体积生物体液中分离出小型细胞外囊泡。
J Extracell Vesicles. 2021 Sep;10(11):e12138. doi: 10.1002/jev2.12138.
9
Density-based lipoprotein depletion improves extracellular vesicle isolation and functional analysis.基于密度的脂蛋白耗竭可改善细胞外囊泡的分离和功能分析。
J Thromb Haemost. 2024 May;22(5):1372-1388. doi: 10.1016/j.jtha.2024.01.010. Epub 2024 Jan 24.
10
Scalable purification of extracellular vesicles with high yield and purity using multimodal flowthrough chromatography.使用多模式流通色谱法以高产量和高纯度可扩展地纯化细胞外囊泡。
J Extracell Biol. 2024 Jan 31;3(2):e138. doi: 10.1002/jex2.138. eCollection 2024 Feb.

引用本文的文献

1
Exploring the therapeutic potential of MSC-derived secretomes in neonatal care: focus on BPD and NEC.探索间充质干细胞分泌产物在新生儿护理中的治疗潜力：聚焦支气管肺发育不良和坏死性小肠结肠炎。
Stem Cell Res Ther. 2025 Aug 29;16(1):476. doi: 10.1186/s13287-025-04616-8.
2
Emerging Biomimetic Drug Delivery Nanoparticles Inspired by Extracellular Vesicles.受细胞外囊泡启发的新型仿生药物递送纳米颗粒
Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2025 Jul-Aug;17(4):e70025. doi: 10.1002/wnan.70025.
3
Emerging biomarkers for early cancer detection and diagnosis: challenges, innovations, and clinical perspectives.

本文引用的文献

1
Challenges in Exosome Isolation and Analysis in Health and Disease.健康与疾病中外泌体分离与分析的挑战。
Int J Mol Sci. 2019 Sep 21;20(19):4684. doi: 10.3390/ijms20194684.
2
The generation and use of recombinant extracellular vesicles as biological reference material.重组细胞外囊泡的产生和应用作为生物参考物质。
Nat Commun. 2019 Jul 23;10(1):3288. doi: 10.1038/s41467-019-11182-0.
3
Mechanisms of intercellular Wnt transport.细胞间 Wnt 运输的机制。
用于早期癌症检测和诊断的新兴生物标志物：挑战、创新与临床前景
Eur J Med Res. 2025 Aug 18;30(1):760. doi: 10.1186/s40001-025-03003-6.
4
The Redox Enzyme Thioredoxin Is Increased in Plasma Extracellular Vesicles From Endurance-Trained Females in Response to Acute Exercise.氧化还原酶硫氧还蛋白在耐力训练女性血浆细胞外囊泡中因急性运动而增加。
FASEB J. 2025 Aug 31;39(16):e70927. doi: 10.1096/fj.202500678R.
5
Proteomic Landscape of Small Extracellular Vesicles Derived From Gastric Juice and Identified TFF2 as a Specific Biomarker.源自胃液的小细胞外囊泡的蛋白质组学特征及鉴定三叶因子2为特异性生物标志物
Int J Nanomedicine. 2025 May 29;20:6929-6948. doi: 10.2147/IJN.S516605. eCollection 2025.
6
Diagnostic and prognostic roles of endothelial- and platelet-derived extracellular vesicles in cardiovascular diseases.内皮细胞和血小板衍生的细胞外囊泡在心血管疾病中的诊断和预后作用。
J Transl Med. 2025 May 16;23(1):553. doi: 10.1186/s12967-025-06522-2.
7
Development of an Effective Isolation Method for Plasma Extracellular Vesicles Excluding Lipoproteins by Precipitation With Polyethylene Glycol.通过聚乙二醇沉淀法开发一种有效的血浆细胞外囊泡分离方法，以排除脂蛋白。
J Extracell Biol. 2025 Apr 25;4(4):e70047. doi: 10.1002/jex2.70047. eCollection 2025 Apr.
8
Circulating lung cancer exosomes damage the niche of intestinal stem cells.循环肺癌外泌体损害肠道干细胞生态位。
Transl Lung Cancer Res. 2025 Mar 31;14(3):718-735. doi: 10.21037/tlcr-24-758. Epub 2025 Mar 10.
9
Optimization of methods for isolation and purification of outer membrane vesicles (OMVs) from Neisseria lactamica.从乳酸奈瑟菌中分离和纯化外膜囊泡（OMV）方法的优化。
Appl Microbiol Biotechnol. 2025 Apr 7;109(1):82. doi: 10.1007/s00253-025-13460-y.
10
The potential of exosomes in regenerative medicine and in the diagnosis and therapies of neurodegenerative diseases and cancer.外泌体在再生医学以及神经退行性疾病和癌症的诊断与治疗中的潜力。
Front Med (Lausanne). 2025 Mar 13;12:1539714. doi: 10.3389/fmed.2025.1539714. eCollection 2025.
Development. 2019 May 15;146(10):dev176073. doi: 10.1242/dev.176073.
4
Comparison of small extracellular vesicles isolated from plasma by ultracentrifugation or size-exclusion chromatography: yield, purity and functional potential.通过超速离心或尺寸排阻色谱法从血浆中分离的小细胞外囊泡的比较：产量、纯度和功能潜力。
J Extracell Vesicles. 2018 Dec 28;8(1):1560809. doi: 10.1080/20013078.2018.1560809. eCollection 2019.
5
Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines.细胞外囊泡研究的最低限度信息2018（MISEV2018）：国际细胞外囊泡协会的立场声明及MISEV2014指南的更新
J Extracell Vesicles. 2018 Nov 23;7(1):1535750. doi: 10.1080/20013078.2018.1535750. eCollection 2018.
6
Specificities of secretion and uptake of exosomes and other extracellular vesicles for cell-to-cell communication.细胞间通讯中外泌体和其他细胞外囊泡的分泌和摄取的特异性。
Nat Cell Biol. 2019 Jan;21(1):9-17. doi: 10.1038/s41556-018-0250-9. Epub 2019 Jan 2.
7
Technical approaches to reduce interference of Fetal calf serum derived RNA in the analysis of extracellular vesicle RNA from cultured cells.减少胎牛血清来源的RNA对培养细胞外泌体RNA分析干扰的技术方法。
J Extracell Vesicles. 2018 Dec 12;8(1):1552059. doi: 10.1080/20013078.2018.1552059. eCollection 2019.
8
Sialic Acid-Dependent Inhibition of T Cells by Exosomal Ganglioside GD3 in Ovarian Tumor Microenvironments.唾液酸依赖性外泌体神经节苷脂 GD3 通过肿瘤微环境抑制 T 细胞。
J Immunol. 2018 Dec 15;201(12):3750-3758. doi: 10.4049/jimmunol.1801041. Epub 2018 Nov 16.
9
Precipitation with polyethylene glycol followed by washing and pelleting by ultracentrifugation enriches extracellular vesicles from tissue culture supernatants in small and large scales.通过聚乙二醇沉淀，随后进行洗涤并通过超速离心进行沉淀，可大规模和小规模地从组织培养上清液中富集细胞外囊泡。
J Extracell Vesicles. 2018 Oct 17;7(1):1528109. doi: 10.1080/20013078.2018.1528109. eCollection 2018.
10
Extracellular vesicles in cancer - implications for future improvements in cancer care.癌症中的细胞外囊泡 - 对未来改善癌症治疗的影响。
Nat Rev Clin Oncol. 2018 Oct;15(10):617-638. doi: 10.1038/s41571-018-0036-9.