基于纳米流式细胞术的细胞外囊泡中线粒体的快速检测

doi:10.5246/jcps.2026.03.021

摘要/Abstract

摘要：

线粒体富集型细胞外囊泡(mitoEVs)作为细胞代谢与组织修复的重要调节者正逐渐受到关注, 但现有的检测方法仍有局限。本研究基于纳米流式细胞术(nano FCM)建立了一套用于检测细胞外囊泡中线粒体的分析方法。通过仪器参数优化, 缓冲液、试剂验证以及简便快速的EV分离流程, 实现了对EV群体中线粒体相关信号的精确检测, 为mitoEV的研究和治疗作用提供了一种可靠且高效的技术手段。

关键词: 线粒体, 细胞外囊泡, 纳米流式细胞术

Abstract:

Mitochondria-rich extracellular vesicles (mitoEVs) are increasingly recognized as key modulators of cellular metabolism and tissue repair, yet their study has been hindered by the lack of sensitive and quantitative detection strategies. Here, we introduced a nanoflow cytometry (nano FCM)–based analytical platform enabling rigorous quantification and phenotypic characterization of mitoEVs. By integrating refined instrument calibration, systematic reagent validation, and a streamlined EV isolation procedure, we achieved reliable detection of mitochondria-associated signatures within defined vesicle populations. This workflow delivered a robust and scalable framework that enhanced mitoEV analysis and supported future applications in mechanistic research and therapeutic development.

Key words: Mitochondria, Extracellular vesicles, Nanoflow cytometry

Supporting:

孙玉芳, 于化龙, 宋书香. 基于纳米流式细胞术的细胞外囊泡中线粒体的快速检测[J]. 中国药学(英文版), 2026, 35(3): 315-323.

Yufang Sun, Hualong Yu, Shuxiang Song. Rapid detection of mitochondria in extracellular vesicles using nanoflow cytometry[J]. Journal of Chinese Pharmaceutical Sciences, 2026, 35(3): 315-323.

图/表 4

Figure 1. (a) Threshold and flow rate; (b) Calibrations of light scatter; (c) Calibrations of fluorescence; (d) Sensitivity monitoring.

Figure 2. (a) PBS only; (b) CD63-PE in PBS; (c) MitoTracker Green in PBS.

Figure 3. (a) EV isolation workflow; (b) EVs from MCF-7 conditioned medium and stained with EV marker CD63; (c) EV from saliva stained with EV marker CD63.

Figure 4. (a) Unstained MCF-7 EV; (b) CD63 single-stained MCF-7 EV; (c) MitoTracker single-stained MCF-7 EV; (d) CD63/MitoTracker double-stained MCF-7 EV; (e) Unstained salivary EV; (f) CD63 single-stained salivary EV; (g) MitoTracker single-stained salivary EV; (h) CD63/MitoTracker double-stained salivary EV.

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