微泡
外体
生物芯片
细胞培养
化学
CD63
溶解
细胞
细胞生物学
癌症研究
生物
生物化学
小RNA
生物信息学
遗传学
基因
作者
Anikó Bertóková,Natalia Svecova,Katarína Kozics,Alena Gábelová,Alica Vikartovská,Eduard Jáné,Michal Hires,Tomáš Bertók,Ján Tkáč
标识
DOI:10.1016/j.biopha.2022.113093
摘要
Exosomes are considered to be a rich source of biomarkers, hence in this article we examine the best procedure for their isolation. We examine several isolation procedures, exosome storage conditions and other conditions affecting exosome production by prostate cell lines. We selected four different commercially available kits based on different principles to achieve exosome isolation, the best being magnetic-based. In addition, we found storage at - 20 °C to be good for storing isolated exosomes and that exosomes were produced from the cancerous prostate cell line 22Rv1 in much greater amounts than the non-cancerous prostate cell line RWPE1. We also found differences in the response of both cell lines in the production of exosomes as a result of stress, i.e. exposure to hydrogen peroxide and starvation. The effect of Triton X-100 on exosome lysis was examined using two different surfactant concentrations by analysis of the exosome count and change in the exosome size. The final part of the article details the advantages of the use of a 2D biochip prepared in-house over a commercially available 3D biochip for monitoring the interaction of exosomes via its surface receptors (CD63) with an immobilised ligand (anti-CD63 antibodies) using surface plasmon resonance. The final experiment shows the potential of lectin fluorescent microarrays for the analysis of glycans present in lysed exosomes.
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