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

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

doi:10.1016/j.xpro.2024.103256

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

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

Research output: Contribution to journal › Article › peer-review

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

Original language	English
Article number	103256
Pages (from-to)	103256
Journal	Star Protocols
Volume	5
Issue number	3
Early online date	Jul 13 2024
DOIs	https://doi.org/10.1016/j.xpro.2024.103256
State	Published - Sep 20 2024

Keywords

Cancer
Cell Biology
Molecular Biology
Animals
Humans
Pancreatic Neoplasms/pathology
Cell Line, Tumor
Mice
Extracellular Vesicles/metabolism
Liver Neoplasms/secondary

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.xpro.2024.103256

Cite this

Casabianca, A., Bellina, M., Dudgeon, C., Nezhad, M. S., Harris, C., Fiore, S., Bernet, A., Narrow, W., Prela, O., Wang, L., Astsaturov, I., Mehlen, P., & Carpizo, D. R. (2024). Protocol for isolating extracellular vesicles from pancreatic cancer cells for liver metastatic niche research. Star Protocols, 5(3), 103256. Article 103256. https://doi.org/10.1016/j.xpro.2024.103256

@article{c68b4b2b21904fec9cb756efe7a1ef1f,

title = "Protocol for isolating extracellular vesicles from pancreatic cancer cells for liver metastatic niche research",

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",

keywords = "Cancer, Cell Biology, Molecular Biology, Animals, Humans, Pancreatic Neoplasms/pathology, Cell Line, Tumor, Mice, Extracellular Vesicles/metabolism, Liver Neoplasms/secondary",

author = "Anthony Casabianca and M{\'e}lanie Bellina and Crissy Dudgeon and Nezhad, {Muhammad Sadeqi} and Chris Harris and Stephany Fiore and Agnes Bernet and Wade Narrow and Orjola Prela and Lan Wang and Igor Astsaturov and Patrick Mehlen and Carpizo, {Darren R.}",

note = "Publisher Copyright: {\textcopyright} 2024 The Authors",

year = "2024",

month = sep,

day = "20",

doi = "10.1016/j.xpro.2024.103256",

language = "English",

volume = "5",

pages = "103256",

journal = "Star Protocols",

issn = "2666-1667",

publisher = "Cell Press",

number = "3",

}

TY - JOUR

T1 - Protocol for isolating extracellular vesicles from pancreatic cancer cells for liver metastatic niche research

AU - Casabianca, Anthony

AU - Bellina, Mélanie

AU - Dudgeon, Crissy

AU - Nezhad, Muhammad Sadeqi

AU - Harris, Chris

AU - Fiore, Stephany

AU - Bernet, Agnes

AU - Narrow, Wade

AU - Prela, Orjola

AU - Wang, Lan

AU - Astsaturov, Igor

AU - Mehlen, Patrick

AU - Carpizo, Darren R.

PY - 2024/9/20

Y1 - 2024/9/20

N2 - 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

AB - 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

KW - Cancer

KW - Cell Biology

KW - Molecular Biology

KW - Animals

KW - Humans

KW - Pancreatic Neoplasms/pathology

KW - Cell Line, Tumor

KW - Mice

KW - Extracellular Vesicles/metabolism

KW - Liver Neoplasms/secondary

UR - http://www.scopus.com/inward/record.url?scp=85201314732&partnerID=8YFLogxK

U2 - 10.1016/j.xpro.2024.103256

DO - 10.1016/j.xpro.2024.103256

M3 - Article

C2 - 39141519

AN - SCOPUS:85201314732

SN - 2666-1667

VL - 5

SP - 103256

JO - Star Protocols

JF - Star Protocols

IS - 3

M1 - 103256

ER -

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

Abstract

Keywords

UN SDGs

Access to Document

Other files and links

Fingerprint

Cite this