4.6 Article

Isolation and analysis of tumor-derived extracellular vesicles from head and neck squamous cell carcinoma plasma by galectin-based glycan recognition particles

期刊

INTERNATIONAL JOURNAL OF ONCOLOGY
卷 61, 期 5, 页码 -

出版社

SPANDIDOS PUBL LTD
DOI: 10.3892/ijo.2022.5423

关键词

extracellular vesicles; exosomes; novel isolation technique; biomarker; head and neck squamous cell carcinoma; tumor-derived extracellular vesicles; galectin-based glycan recognition particles; beads-based flow cytometry

类别

向作者/读者索取更多资源

This study investigated the practicality of galectin-based magnetic glycan recognition particles (EXOBead) as a novel EV isolation technique for squamous cell carcinoma of the head and neck. The results showed high concentration of pure EVs and detection of specific EV markers, indicating low contamination compared with size exclusion chromatography. Bead-based flow cytometry provided an easy way to understand EV subpopulations. Overall, EXOBead showed great potential as a new isolation tool with high throughput capacity that could potentially be used in a clinical setting.
Extracellular vesicles (EVs) have recently come into the spotlight as potential cancer biomarkers. Isolation of pure EVs is complex, so wider use requires reliable and time-efficient isolation methods. In the present study, galectin-based magnetic glycan recognition particles, EXOBead (R) were investigated for their practicality as a novel EV isolation technique, exemplified here for squamous cell carcinoma of the head and neck. Analysis of the isolation method showed a high concentration of pure EVs with detection of specific EV markers such as CD9, CD63, CD81 and TSG101. No apolipoprotein A1 was shown in the isolates, indicating low contamination of this isolation technique compared with size exclusion chromatography. In addition, common leukocyte antigen (CD45), three HNSCC [epithelial cell adhesion molecule (EpCAM), pan-cytokeratin and programmed death-ligand 1 (PD-L1)] and PanEV markers (premixed CD9, CD63 and CD81 antibodies) were measured by bead-based flow cytometry (BFC). BFC revealed that CD45(Neg) PanEV(+), EpCAM(+) PanEV(+) and PD-L1(+) PanEV(+) were significantly higher in tumor patients compared with healthy control plasma. CD45(Neg) PanEV(+) and CD45(+) PanEV(+) carrying two or three HNSCC biomarkers were also significantly higher in tumor patients compared with healthy controls (BFC). Comparison of the functional immunosuppression effect of eluted tumor patient plasma EVs from EXOBead (R) and commercial polyethylene glycol isolation showed a significant tumor-dependent increase in concentration of EVs. A peripheral blood mononuclear cell activation assay also showed that the T-cell functionality of tumor patient plasma EVs isolated with EXOBead (R) was preserved in vitro. In conclusion, isolation using galectin-based magnetic glycan recognition particles is a novel method for isolating plasma EVs with low lipoprotein contamination. Bead-based flow cytometry provided an easy way to understand EV subpopulations. EXOBead (R) therefore showed great potential as a new isolation tool with high throughput capacity that could potentially be used in a clinical setting.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.6
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据