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改良的非小细胞肺癌细胞(NC1975)外泌体分离方法及其使用形态学和表面蛋白生物标志物方法进行的表征。

Improved exosome isolation methods from non-small lung cancer cells (NC1975) and their characterization using morphological and surface protein biomarker methods.

机构信息

Science and Engineering Department, Hamad Bin Khalifa University, P. O. Box 34110, Doha, Qatar.

Mechanical and Industrial Engineering Department, Qatar University, P. O. Box 2713, Doha, Qatar.

出版信息

J Cancer Res Clin Oncol. 2023 Aug;149(10):7505-7514. doi: 10.1007/s00432-023-04682-6. Epub 2023 Mar 25.

Abstract

This study has demonstrated improved methods for isolating exosomes from non-small lung cancer cells, which address the problems characterized by exosome morphological and chemical methods. To improve the isolation methods, cells from the NCI 1975 cell line were used as the source for exosomes. The isolation processes were carried out using serial isolation techniques in addition to specific preservation tools. The isolated exosomes were characterized using transmission electron microscopy (TEM), and scanning electron microscopy (SEM) was added for further assurance of the investigation results. The statistical analysis results showed that the size distributions of apoptotic vesicles (APV) 450 nm and necrotic bodies (NCB) 280 nm (extracellular vesicles) were significantly different from exosomes (P < 0.001). In contrast, the exosome size distribution was not significantly different from the published exosome sizes, as demonstrated by statistical analysis tools. This study confirmed the improved methods for isolating exosomes that make exosomes accessible for use in the diagnosis and prognosis of non-small cell lung cancer (NSCLC).

摘要

本研究展示了改良的非小细胞肺癌细胞外泌体分离方法,解决了外泌体形态学和化学方法所具有的问题。为了改进分离方法,我们使用 NCI 1975 细胞系作为外泌体的来源。采用连续分离技术,并结合特定的保存工具进行分离过程。使用透射电子显微镜(TEM)对分离出的外泌体进行特征描述,并通过扫描电子显微镜(SEM)进一步确保研究结果。统计分析结果表明,凋亡小体(APV)450nm 和坏死体(NCB)280nm(细胞外囊泡)的大小分布与外泌体有显著差异(P<0.001)。相比之下,外泌体的大小分布与通过统计分析工具所展示的已发表的外泌体大小没有显著差异。本研究证实了改良的外泌体分离方法可用于非小细胞肺癌(NSCLC)的诊断和预后。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/718c/10374817/0d7c9ede2fe2/432_2023_4682_Fig1_HTML.jpg

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