Isolation and Analysis of Tumor-Derived Exosomes.


Journal

Current protocols in immunology
ISSN: 1934-368X
Titre abrégé: Curr Protoc Immunol
Pays: United States
ID NLM: 9101651

Informations de publication

Date de publication:
12 2019
Historique:
entrez: 26 11 2019
pubmed: 26 11 2019
medline: 8 7 2020
Statut: ppublish

Résumé

A method for isolation of exosomes from tumor cell supernatants or cancer patients' plasma is presented. Tumor-derived exosomes (TEX) are defined as a subset of extracellular vesicles (EVs) sized at 30 to 150 nm and originating from multivesicular bodies (MVBs). The method utilizes size exclusion chromatography (SEC) for recovery of exosomes from cell-line supernatants or cancer patients' plasma. The recovered exosomes are morphologically intact, aggregate-free, and functionally competent. Their molecular content parallels that of the parent tumor cells and they carry various immunoregulatory ligands known to modulate functions of immune cells. All exosomes isolated from tumor cell lines are TEX, while those isolated from plasma of cancer patients have to be fractionated into TEX and non-TEX. Mini-SEC allows for exosome isolation and recovery in quantities sufficient for molecular profiling, functional studies, and, in the case of plasma, further fractionation into TEX and non-TEX. The mini-SEC method can also be used for comparative studies of the exosome content in serial specimens of cancer patients' body fluids. © 2019 by John Wiley & Sons, Inc.

Identifiants

pubmed: 31763776
doi: 10.1002/cpim.91
pmc: PMC6880756
mid: NIHMS1046334
doi:

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

e91

Subventions

Organisme : NCI NIH HHS
ID : R01 CA168628
Pays : United States
Organisme : NCI NIH HHS
ID : R21 CA205644
Pays : United States

Informations de copyright

© 2019 John Wiley & Sons, Inc.

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Auteurs

Nils Ludwig (N)

Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.
UPMC Hillman Cancer Center, Pittsburgh, Pennsylvania.

Chang-Sook Hong (CS)

Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.
UPMC Hillman Cancer Center, Pittsburgh, Pennsylvania.

Sonja Ludwig (S)

Department of Otolaryngology, University of Duisburg-Essen, Essen, Germany.

Juliana H Azambuja (JH)

Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.
UPMC Hillman Cancer Center, Pittsburgh, Pennsylvania.
Programa de Pós-Graduação em Biociências, Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA), Porto Alegre, Brazil.

Priyanka Sharma (P)

Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.
UPMC Hillman Cancer Center, Pittsburgh, Pennsylvania.

Marie-Nicole Theodoraki (MN)

Department of Otorhinolaryngology, Head and Neck Surgery, University of Ulm, Ulm, Germany.

Theresa L Whiteside (TL)

Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.
UPMC Hillman Cancer Center, Pittsburgh, Pennsylvania.
Department of Immunology, University of Pittsburgh, Pittsburgh, Pennsylvania.
Department of Otolaryngology, University of Pittsburgh, Pittsburgh, Pennsylvania.

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