单细胞与单颗粒外泌体分离分析进展与展望  

Progress and prospect of separation and analysis of single-cell and single-particle exosomes

作　　者：卜爱香 武光耀 胡良海[1] BU Aixiang;WU Guangyao;HU Lianghai(Center for Supramolecular Chemical Biology,State Key Laboratory of Supramolecular Structure and Materials,Jilin University,Changchun 130023,China)

机构地区：[1]吉林大学超分子化学生物学研究中心,超分子结构与材料全国重点实验室,生命科学学院,吉林长春130023

出　　处：《色谱》2025年第5期399-412,共14页Chinese Journal of Chromatography

基　　金：国家自然科学基金(22374056).

摘　　要：外泌体是细胞分泌的一种纳米级囊泡颗粒,由脂质双分子层包裹,它们在细胞通讯中发挥着重要的作用,并参与到多种生理及病理过程中,如免疫调节、血管生成以及肿瘤的发生与转移等。外泌体携带了来自母体细胞的多种生物分子,因此成为疾病标志物发现的一个重要载体。由于来自不同细胞亚群的外泌体具有显著的异质性,利用传统检测方法仅能获取样本中群体细胞的平均信息,而无法建立外泌体生物功能与亚型之间的明确关联。因此,为了深入探究外泌体的特异性并清晰区分不同亚型外泌体的特征,有必要在单细胞及单颗粒水平上对外泌体进行表征。常用的单颗粒外泌体表征技术主要包括流式细胞术、超分辨显微镜、原子力显微镜、表面增强拉曼光谱、邻近编码技术和质谱技术等。本文总结了基于微流控技术的单细胞外泌体分离分析方法及单颗粒外泌体表征技术的最新进展,并对新兴技术(如Olink蛋白质组学、点击化学和分子印迹技术等)在单细胞及单颗粒外泌体研究中的应用前景进行了展望。Exosomes are nanoscale vesicles secreted by cells and are encapsulated in lipid bilayers.They play crucial roles in cell communication and are involved in a variety of physiological and pathological processes,including immune regulation,angiogenesis,and tumor initiation and metastasis.Exosomes carry a variety of biomolecules from maternal cells and are therefore important vehicles for discovering disease markers.Traditional detection methods only provide average cell-population information for a given sample and cannot establish clear relationships between the biological functions of exosomes and subtype owing to the significant heterogeneity associated with exosomes from different cell subsets.Therefore,characterizing exosomes at the single-cell and single-particle levels requires exosome specificities to be further explored and the characteristics of various exosome subtypes to be distinguished.Commonly used single-particle exosome characterization technologies include flow cytometry,super-resolution microscopy,atomic force microscopy,surface-enhanced Raman spectroscopy,proximity barcoding assay and MS.In this paper,we summarize recent advances in the separation and characterization of single-cell exosomes based on microfluidics and provide future applications prospects for emerging technologies(such as Olink proteomics,click chemistry,and molecular imprinting)for studying single-cell and single-particle exosomes.

关 键 词：单细胞外泌体 单颗粒外泌体 分离与分析 微流控技术 综述 

分 类 号：O658[理学—分析化学]