Low-power sonication can reduce the size of SHED-MSCs-derived exosomes
Low-power sonication can reduce the size of SHED-MSCs-derived exosomes
Azadeh Mohammad-Hasani,1Ali Fallah,2Ayyoob Khosravi,3,*Mohsen Saeidi,4Saeed Mohammadi,5
1. Department of Molecular Medicine, Faculty of Advanced Medical Technologies, Golestan University of Medical Sciences, Gorgan, Iran 2. Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar 3. Department of Molecular Medicine, Faculty of Advanced Medical Technologies, Golestan University of Medical Sciences, Gorgan, Iran 4. Department of Immunology, School of Medicine, Golestan University of Medical Sciences, Gorgan, Iran 5. Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, Iran, Stem Cell Research Center, Golestan University of Medical Sciences, Gorgan, Iran
Introduction: Exosomes isolated under laboratory conditions are prone to aggregation, which causes problems in identifying their characteristics such as particle size and concentration. This study investigated the effects of sonication at different time periods and then analyzed the properties of SHED-MSCs-derived exosomes using DLS and AFM techniques.
Methods: After incubating the SHED-MSCs in a serum-free medium, the supernatant was collected and SHED-MSCs-Exo was isolated using the Exocib Exosome Isolation Kit (Cibbiotech, Tehran, Iran) according to the manufacturer's protocol. The extracted exosomes were divided into three groups: sonication, 5 min sonication, and 10 min sonication.
Results: DLS and AFM analyses of the three groups of exosomes show that the size of exosomes before sonication is in the range of 100-1000 nm, while after 5 min of sonication, their size is in two ranges or two peaks of 10-100 nm and 100-1000 nm. In the third group, after 10 minutes of sonication, all exosomes were found to be between 10 - 100 nm in size.
Conclusion: Low-power sonication has been widely used to dissociate aggregates of isolated exosomes prior to analysis. This technique separates particles by vibrations in a suspension.