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开发一种用于即时检测环境中细胞外囊泡分离的微流控装置。

Development of a Microfluidic Device for Exosome Isolation in Point-of-Care Settings.

机构信息

Asghar-Lab-Micro and Nanotechnology in Medicine Lab, Florida Atlantic University, Boca Raton, FL 33431, USA.

Department of Biological Sciences, Florida Atlantic University, Boca Raton, FL 33431, USA.

出版信息

Sensors (Basel). 2023 Oct 7;23(19):8292. doi: 10.3390/s23198292.

Abstract

Exosomes have gained recognition in cancer diagnostics and therapeutics. However, most exosome isolation methods are time-consuming, costly, and require bulky equipment, rendering them unsuitable for point-of-care (POC) settings. Microfluidics can be the key to solving these challenges. Here, we present a double filtration microfluidic device that can rapidly isolate exosomes via size-exclusion principles in POC settings. The device can efficiently isolate exosomes from 50-100 µL of plasma within 50 min. The device was compared against an already established exosome isolation method, polyethylene glycol (PEG)-based precipitation. The findings showed that both methods yield comparable exosome sizes and purity; however, exosomes isolated from the device exhibited an earlier miRNA detection compared to exosomes obtained from the PEG-based isolation. A comparative analysis of exosomes collected from membrane filters with 15 nm and 30 nm pore sizes showed a similarity in exosome size and miRNA detection, with significantly increased sample purity. Finally, TEM images were taken to analyze how the developed devices and PEG-based isolation alter exosome morphology and to analyze exosome sizes. This developed microfluidic device is cost-efficient and time-efficient. Thus, it is ideal for use in low-resourced and POC settings to aid in cancer and disease diagnostics and therapeutics.

摘要

外泌体在癌症诊断和治疗中得到了认可。然而,大多数外泌体分离方法耗时、昂贵,且需要大型设备,因此不适合即时检测(POC)环境。微流控技术可能是解决这些挑战的关键。在这里,我们提出了一种双重过滤微流控装置,可通过尺寸排阻原理在 POC 环境中快速分离外泌体。该装置可在 50 分钟内从 50-100μL 的血浆中有效分离外泌体。该装置与已建立的外泌体分离方法——聚乙二醇(PEG)沉淀进行了比较。结果表明,两种方法得到的外泌体大小和纯度相当;然而,与从 PEG 分离得到的外泌体相比,从该装置分离得到的外泌体更早检测到 miRNA。对 15nm 和 30nm 孔径的膜过滤器收集的外泌体进行比较分析表明,外泌体大小和 miRNA 检测具有相似性,且样品纯度显著提高。最后,通过 TEM 图像分析了开发的设备和基于 PEG 的分离如何改变外泌体的形态并分析外泌体的大小。这种开发的微流控装置具有成本效益和时间效益。因此,它非常适合在资源有限和 POC 环境中使用,以辅助癌症和疾病的诊断和治疗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6aae/10574868/88c6dee918fe/sensors-23-08292-g001.jpg

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