Protocol for isolating extracellular vesicles from pancreatic cancer cells for liver metastatic niche research

从胰腺癌细胞中分离细胞外囊泡以用于肝转移性微环境研究的方案

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作者:Anthony Casabianca, Mélanie Bellina, Crissy Dudgeon, Muhammad Sadeqi Nezhad, Chris Harris, Stephany Fiore, Agnes Bernet, Wade Narrow, Orjola Prela, Lan Wang, Igor Astsaturov, Patrick Mehlen, Darren R Carpizo

Abstract

Extracellular vesicles (EVs) are secreted, cell-derived, membrane-bound compartments implicated in various diseases for their ability to influence distant targets and as carriers of biomarkers. Here, we present a protocol for separating EVs from mammalian pancreatic cancer cells and their characterization using western blot and electron microscopy. We then demonstrate how they are utilized to affect tumor development in a murine model of metastatic pancreatic cancer including a method to quantify hepatic tumor burden in histologic samples. For complete details on the use and execution of this protocol, please refer to Dudgeon et al.1.

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