可同时浸渍伊马替尼的外泌体分离微芯片:体外分析

Pub Date : 2024-03-25 DOI:10.15275/rusomj.2024.0104
Amir Monfaredan, Fakher Rahim, Gholamreza Tavoosidana, M. Modarressi, Alaviyehsadat Hosseininasab, Ali-Akbar Aghajani-Afrouzi, Mahdi Shafiee Sabet, E. Motevaseli
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引用次数: 0

摘要

背景与目的--外泌体是一种源自真核细胞的微小双层膜,因其尺寸适中、与生物体相容、结构坚固、可携带大量货物以及表面可修饰而被认为是一种宝贵的天然载体。方法 - 由于保持外泌体的高纯度所面临的挑战,人们采用了各种策略来分离外泌体。目前的研究利用软光刻技术制造了用于分离外泌体的通道,并结合了免疫亲和功能。进行了生化和生物物理检测,以评估从不同来源(血清、细胞上清液和尿液)分离的外泌体的质量,并与市售试剂盒进行比较。结果 - 目前的研究采用微流控方法捕获 CD63 结合物磁珠,从而非常有效地分离了外泌体。根据数据,miRNA 没有明显的统计学差异。这表明工程芯片成功实现了外泌体的分离,同时保持了其核酸成分的完整性。结论--研究结果表明,目前的方法能有效分离外泌体,而且产量高、纯度高、所需时间短。含有伊马替尼的外泌体对 KYO-1 细胞系的各种形式的癌症均有抗癌效果。
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A Microchip For Exosome Isolation That Can Be Impregnated With Imatinib Simultaneously: An In Vitro Analysis
Background and Aims — Exosomes, which are tiny double-layered membranes originating from eukaryotic cells, have been recognized as a valuable natural vehicle for delivering substances because of their optimal size, compatibility with living organisms, strong structure, ability to carry a large amount of cargo, and capacity to be modified on their surface. Methods — Various strategies have been employed to isolate exosomes due to the challenges associated with maintaining their high purity. The current investigation utilized a soft lithography technique to fabricate channels for exosome separation, incorporating immunoaffinity capabilities. Both biochemical and biophysical assays were conducted to assess the quality of isolated exosomes from various sources (serum, cell supernatant, and urine) and compared with a commercially available kit. Results — The current investigation employed a microfluidic method to capture CD63-conjugated magnetic beads, resulting in a very effective separation of exosomes. Based on the data, there were no notable variations in miRNAs that were statistically significant. This demonstrates that the engineered chip successfully achieved the separation of the exosome while preserving the integrity of its nucleic acid components. Conclusion — The results shown that the current methodology effectively isolated exosomes with a high yield rate, purity, and minimal time requirement. The imatinib laden exosomes demonstrated anticancer efficacy against the KYO-1 cell line in all of their forms.
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