JOVE publishes a series of exosome technique videos

Isolation and Characterization of RNA-Containing Exosomes

Cecilia Lässer*¹, Maria Eldh*¹, Jan Lötvall¹

¹Krefting Research Centre, Department of Internal Medicine, Sahlgrenska Academy, University of Gothenburg

This paper demonstrates methods for the isolation, purification and detection of exosomes, as well as techniques for analysis of their molecular content. These methods are adaptable for exosome isolation from both cell culture media and biological fluids, and can beyond analysis of molecular content also be useful in functional studies.

Published January 9, 2012. Keywords: Molecular Biology, Exosomes, microvesicles, mRNA, miRNA, RNA isolation, flow cytometry, electron microscopy, Western blot, Bioanalyzer

A Modified Precipitation Method to Isolate Urinary Exosomes

Rupesh Kanchi Ravi¹, Mahdieh Khosroheidari¹, Johanna K. DiStefano¹

¹Diabetes, Cardiovascular and Metabolic Diseases Division, Translational Genomics Research Institute (TGen)

This manuscript describes a protocol for isolating exosomes from human urine using a modified precipitation technique.

Published January 16, 2015. Keywords: Medicine, Translational medicine, exosomes, urine, RNA, western blot, Tamm-Horsfall Protein

Detection of Exosomal Biomarker by Electric Field-induced Release and Measurement (EFIRM)

Michael Tu¹, Fang Wei¹, Jieping Yang², David Wong¹

¹School of Dentistry, University of California, Los Angeles, ²School of Medicine, Clinical Nutrition, University of California, Los Angeles

Exosomes are microvesicular structures found within biofluids that potentially carry important disease discriminatory biomarkers. Here, a novel method is used to specifically extract exosomes and rapidly test the exosomal cargo for both RNA/protein targets following the disruption of exosomes using non-uniform electric cyclic square waves.

Published January 23, 2015. Keywords: Bioengineering, Exosome, Electrochemical sensors, Tumor biomarkers, Lung cancer, Salivary diagnostics

An Innovative Method for Exosome Quantification and Size Measurement

Arash Mehdiani¹, Anatol Maier¹, Antonio Pinto¹, Mareike Barth¹, Payam Akhyari¹, Artur Lichtenberg¹

¹Research Group Experimental Surgery, Department of Cardiovascular Surgery, Medical Faculty, Heinrich-Heine-University Dusseldorf

A method for isolation of exosomes from whole blood and further analysis by nanoparticle tracking using a semi-automatic instrument is presented in this article. The presented technology provides an extremely sensitive method for visualizing and analyzing particles in liquid suspension.

Published January 17, 2015. Keywords: Biochemistry, Exosome, exosome separation, microvesicle, nanoparticle-tracking analysis, nanoparticle counting, particle size analysis, ultracentrifugation, exosome isolation, blood

Quantification and Size-profiling of Extracellular Vesicles Using Tunable Resistive Pulse Sensing

Sybren L. N. Maas^1,2, Jeroen De Vrij^1,2, Marike L. D. Broekman^1,2

¹Department of Neurosurgery, University Medical Center Utrecht, ²Brain Center Rudolf Magnus, University Medical Center Utrecht

Extracellular vesicles play important roles in physiological and pathological processes, including coagulation, immune responses, and cancer or as potential therapeutic agents in drug delivery or regenerative medicine. This protocol presents methods for the quantification and size characterization of isolated and non-isolated extracellular vesicles in various fluids using tunable resistive pulse sensing.

Published October 19, 2014. Keywords: Bioengineering, exosomes, microvesicles, extracellular vesicles, quantification, characterization, Tunable Resistive Pulse Sensing, qNano

Purification and microRNA Profiling of Exosomes Derived from Blood and Culture Media

Marguerite K. McDonald¹, Kathryn E. Capasso¹, Seena K. Ajit¹

¹Department of Pharmacology & Physiology, Drexel University College of Medicine

The presence of stable microRNAs (miRNAs) in exosomes has generated immense interest as a novel mode of intercellular communication, for their potential utility as biomarkers and as a route for therapeutic intervention. Here we demonstrate exosome purification from blood and culture media followed by quantitative PCR to identify miRNAs being transported.

Published June 14, 2013. Keywords: Genetics, Molecular Biology, Cellular Biology, Medicine, Biochemistry, Genomics, Pharmacology, Exosomes, RNA, MicroRNAs, Biomarkers, Pharmacological, Exosomes, microRNA, qPCR, PCR, blood, biomarker, TLDA, profiling, sequencing, cell culture