Protocol for isolating patient-derived ascites cells and extracellular vesicles from gastric cancer peritoneal metastases
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.<sup>1</sup>.
Authors
- George A. Călin (ORCID: https://orcid.org/0000-0002-7427-0578)
- Rebecca Waters (ORCID: https://orcid.org/0000-0002-8163-7497)
- Yibo Fan (ORCID: https://orcid.org/0000-0001-7589-4158)
- Melissa Pool Pizzi (ORCID: https://orcid.org/0000-0002-2373-0052)
- Gengyi Zou (ORCID: https://orcid.org/0000-0002-5399-8674)
- Qiong Gan (ORCID: https://orcid.org/0000-0001-6563-5611)
- Samir Hanash (ORCID: https://orcid.org/0000-0002-4210-1593)
- Jody V. Vykoukal (ORCID: https://orcid.org/0000-0001-7797-627X)
- Hiroyuki Katayama (ORCID: https://orcid.org/0000-0003-3117-1390)
- Linghua Wang (ORCID: https://orcid.org/0000-0001-9380-0266)
- Shilpa S. Dhar (ORCID: https://orcid.org/0000-0002-5922-2164)
- Jiankang Jin (ORCID: https://orcid.org/0000-0001-9864-9817)
- J. A. Ajani
- Shumei Song
Institutions
- The University of Texas MD Anderson Cancer Center (US)
- Coriell Institute For Medical Research (US)
- Hamamatsu University School of Medicine (JP)
- Hamamatsu Medical Center (JP)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-10-17
- DOI
- https://doi.org/10.1016/j.xpro.2025.104133
- Primary Topic
- Extracellular vesicles in disease
- Type
- article
- Field-Weighted Citation Impact
- 0.00