Isolation of extracellular vesicles from intestinal tissue in a mouse model of intestinal ischemia/reperfusion injury | Zendy

Xiao-Dong Chen | Zendy; Jin Zhao | Zendy; Zhengzheng Yan | Zendy; Bowei Zhou | Zendy; Wenfang Huang | Zendy; Wei-Feng Liu | Zendy; Cai Li | Zendy; KeXuan Liu | Zendy

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Isolation of extracellular vesicles from intestinal tissue in a mouse model of intestinal ischemia/reperfusion injury

Author(s) -

Xiao-Dong Chen,

Jin Zhao,

Zhengzheng Yan,

Bowei Zhou,

Wenfang Huang,

Wei-Feng Liu,

Cai Li,

KeXuan Liu

Publication year - 2020

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/btn-2019-0159

Subject(s) - microvesicles , cd63 , microbiology and biotechnology , extracellular vesicles , reperfusion injury , microrna , exosome , intestinal ischemia , extracellular vesicle , intracellular , extracellular , chemistry , ischemia , biology , biochemistry , medicine , gene , cardiology

Extracellular vesicles (EVs) are small membranous particles that contribute to intercellular communications. Separating EVs from tissue is still a technical challenge. Here, we present a rigorous method for extracting EVs from intestinal tissue in a mouse intestinal ischemia/reperfusion (I/R) model, and for analyzing their miRNA content. The isolated EVs show a typical cup shape with a size peak of 120–130 nm in diameter, confirmed by TEM and NTA. They also express EV markers such as CD9, CD63, CD81, Tsg101 and Alix. Real-time qPCR confirmed that these pellets contain miRNAs related to I/R injury. Our study presents a practical way to isolate EVs from intestinal tissue which is suitable for downstream applications such as miRNA analysis, and provides a novel method for investigating the mechanism of intestinal I/R injury.

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