Abstract
This chapter describes a method for isolating human salivary extracellular vesicles (EVs) using density gradient ultracentrifugation. Standard protocols established for isolation of EVs from blood or a conditioned medium of cultured cells do not work for whole saliva, due to its viscosity. Therefore, procedures including a pretreatment step and utilizing iodixanol as a gradient material enable EVs to be concentrated to a 1.1 g/ml density. This protocol is compatible with both swing and angle rotors. By employing an angle rotor, which enables high g-force, the centrifugation time was reduced to 4 h from the 17 h required when using a swing rotor.
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Acknowledgments
This work was supported by the “Project for Private Universities: Matching Fund Subsidy” from the Ministry of Education, Culture, Sports, Science, and Technology and by a grant from the Vehicle Racing Commemorative Foundation.
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Iwai, K., Yamamoto, S., Yoshida, M., Shiba, K. (2017). Isolation of Extracellular Vesicles in Saliva Using Density Gradient Ultracentrifugation. In: Kuo, W., Jia, S. (eds) Extracellular Vesicles. Methods in Molecular Biology, vol 1660. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7253-1_27
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DOI: https://doi.org/10.1007/978-1-4939-7253-1_27
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