Document Type : Research Articles
Authors
1
Division of Biotechnology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran
2
Division of Biotechnology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran
10.22067/jcmr.2026.98095.1132
Abstract
Exosomes are extracellular nanovesicles secreted by almost all cells and are present in all body fluids. These natural carriers, which transport biological molecules, consist of proteins, nucleic acids, and lipids. Exosomes' composition reflects the nature and physiological state of their producer cells and plays a widespread role from intercellular communication in normal physiological conditions to contributing to disease progression. There has been increased interest in exosomes as a novel class of biomarkers for diagnosing various diseases or as delivery vehicles. Therefore, the development of an effective exosome isolation method is essential. Some isolation techniques, such as differential ultracentrifugation, ultrafiltration, and size-exclusion chromatography, face challenges in exosome isolation, such as disruption of exosome membrane integrity and loss of exosome functionality, high operational cost, or time-consuming procedure. In this study, we present a polyethylene glycol (PEG)-based method that offers a simple, cost-effective, and efficient way to isolate exosomes from cell culture supernatants. This method aims to overcome limitations associated with conventional isolation techniques, such as high cost and technical complexity. Here, we aimed to optimize a PEG-based method to improve exosome recovery while preserving vesicle integrity. To achieve this, CD9-stably expressing cells were constructed to enhance exosome production. By evaluating the size and morphology of the isolated exosomes using dynamic light scattering (DLS) and transmission electron microscopy, we demonstrated that washing PEG-enriched exosomes with phosphate-buffered saline followed by centrifugation effectively removed excess PEG, yielding exosomes with a Z-average diameter of 170 nm and a polydispersity index of 0.2, indicating natural and uniform particle size.
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