Purification of leukemia-derived exosomes to study microenvironment modulation

Marina Wierz, Sandrine Pierson, Ernesto Gargiulo, Coralie Guerin, Etienne Moussay, Jerome Paggetti*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

7 Citations (Scopus)

Abstract

Exosomes are membrane-enclosed vesicles released by different cell types into the extracellular space. As mediators of intercellular communication, they are involved in multiple physiological processes, but they are also associated with the pathogenesis of human malignancies including leukemia. Isolation of exosomes enables the characterization of their role in microenvironment modulation as well as their participation in disease pathology. A variety of strategies and techniques exists to purify exosomes from many biological fluids (e.g., blood, urine, and saliva). Here, we describe the efficient production of large quantities of exosomes from leukemic cell lines by using CELLine bioreactors based on two-compartment technology, as well as their isolation and purification by combining differential centrifugation and ultracentrifugation through a density gradient (17% OptiPrep™ cushion). Thus, exosomes are appropriately prepared for characterization by western blotting to detect exosome markers or imaging flow cytometry (ImageStream), and for downstream analyses such as the internalization in microenvironmental cells by confocal imaging or flow cytometry, methods which are also described in this chapter.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages231-245
Number of pages15
DOIs
Publication statusPublished - 2019

Publication series

NameMethods in Molecular Biology
Volume1884
ISSN (Print)1064-3745

Keywords

  • Density gradient ultracentrifugation
  • Exosomes
  • Imaging flow cytometry
  • Intercellular communication
  • Leukemia
  • Purification
  • Two-compartment bioreactor

Fingerprint

Dive into the research topics of 'Purification of leukemia-derived exosomes to study microenvironment modulation'. Together they form a unique fingerprint.

Cite this