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Protocol for isolating patient-derived ascites cells and extracellular vesicles from gastric cancer peritoneal metastases.

STAR protocols 2025 Vol.6(4) p. 104133

Fan Y, Vykoukal JV, Song S, Pizzi MP, Zou G, Yoshimura K, Jin J, Katayama H, Calin GA, Waters RE, Gan Q, Wang L, Hanash S, Dhar SS, Ajani JA

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Peritoneal carcinomatosis (PC) cells and extracellular vesicles (EVs) from gastric cancer ascites are valuable for studying tumor-stroma interactions.

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APA Fan Y, Vykoukal JV, et al. (2025). Protocol for isolating patient-derived ascites cells and extracellular vesicles from gastric cancer peritoneal metastases.. STAR protocols, 6(4), 104133. https://doi.org/10.1016/j.xpro.2025.104133
MLA Fan Y, et al.. "Protocol for isolating patient-derived ascites cells and extracellular vesicles from gastric cancer peritoneal metastases.." STAR protocols, vol. 6, no. 4, 2025, pp. 104133.
PMID 41105517

Abstract

Peritoneal carcinomatosis (PC) cells and extracellular vesicles (EVs) from gastric cancer ascites are valuable for studying tumor-stroma interactions. Here, we present a protocol for isolating PC cells and EVs from patient ascites. We describe steps for PC cell culture, iodixanol-based EV purification, electron microscopy, nanoparticle tracking analysis, flow cytometry, and proteomic profiling. This protocol also includes TD-139 loading into exosomes for functional assays to evaluate their role in modulating the tumor microenvironment. For complete details on the use and execution of this protocol, please refer to Fan et al..

MeSH Terms

Humans; Stomach Neoplasms; Extracellular Vesicles; Ascites; Peritoneal Neoplasms; Proteomics; Tumor Microenvironment; Flow Cytometry; Cell Line, Tumor; Cell Separation

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