Using Imaging Flow Cytometry to Characterize Extracellular Vesicles Isolated from Cell Culture Media, Plasma or Urine

John R Woollard; Amrutesh Puranik; Kyra L Jordan; Lilach O Lerman

doi:10.21769/BioProtoc.3420

Using Imaging Flow Cytometry to Characterize Extracellular Vesicles Isolated from Cell Culture Media, Plasma or Urine

Bio Protoc. 2019 Nov 5;9(21):e3420. doi: 10.21769/BioProtoc.3420.

Authors

John R Woollard¹, Amrutesh Puranik², Kyra L Jordan¹, Lilach O Lerman¹

Affiliations

¹ Division of Nephrology & Hypertension Mayo Clinic, Rochester, MN, United States.
² Division of Rheumatology, Department of Medicine, New York University Langone Medical Center, New York, NY, United States.

Abstract

The ability to non-invasively detect specific damage to the kidney has been limited. Identification of extracellular vesicles released by cells, especially when under duress, might allow for monitoring and identification of specific cell types within the kidney that are stressed. We have adapted a previously published traditional flow cytometry method for use with an imaging flow cytometer (Amnis FlowSight) for identifying EV released by specific cell types and excreted into the urine or blood using markers characteristic of particular cells in the kidney. Here we present a protocol utilizing the Amnis FlowSight Imaging Flow Cytometer to identify and quantify EV from the urine of patients with essential hypertension and renovascular disease. Notably, EV isolated from cell culture media and plasma can also be analyzed similarly.

Keywords: Biomarkers; Exosomes; Extracellular vesicles; Hypertension; Imaging flow cytometry; Renovascular disease.