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Methods Mol Biol. 2017;1660:343-350. doi: 10.1007/978-1-4939-7253-1_27.

Isolation of Extracellular Vesicles in Saliva Using Density Gradient Ultracentrifugation.

Author information

1
Division of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, Tokyo, Japan.
2
Department of Oral & Maxillofacial Implantology, Tokyo Dental College, Tokyo, Japan.
3
Division of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, Tokyo, Japan. kshiba@jfcr.or.jp.

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.

KEYWORDS:

Angle rotor; EVs; Exosome; Extracellular vesicles; Iodixanol; Whole saliva

PMID:
28828669
DOI:
10.1007/978-1-4939-7253-1_27
[Indexed for MEDLINE]

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