Isolation of Extracellular Vesicles in Saliva Using Density Gradient Ultracentrifugation

Kazuya Iwai; Satoshi Yamamoto; Mitsutaka Yoshida; Kiyotaka Shiba

doi:10.1007/978-1-4939-7253-1_27

Isolation of Extracellular Vesicles in Saliva Using Density Gradient Ultracentrifugation

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

Authors

Kazuya Iwai^{1

2}, Satoshi Yamamoto^{1

2}, Mitsutaka Yoshida^{1

2}, Kiyotaka Shiba³

Affiliations

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.

PMID: 28828669
DOI: 10.1007/978-1-4939-7253-1_27

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Centrifugation, Density Gradient*
Extracellular Vesicles / metabolism*
Humans
Saliva / metabolism*
Ultracentrifugation*
Workflow