Journal
FRONTIERS IN BIOSCIENCE-LANDMARK
Volume 23, Issue -, Pages 865-874Publisher
FRONTIERS IN BIOSCIENCE INC
DOI: 10.2741/4621
Keywords
Extracellular vesicles; Plasma; Exosomes; Ultracentrifugation; Size exclusion chromatography; Bovine
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Funding
- New Zealand Ministry of Business, Innovation and Employment [DRCX1302]
- New Zealand dairy farmers through DairyNZ Inc. [DRCX1302]
- Shimadzu Corporation
- Australian Research Council
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Exosomes are a subset of extracellular vesicles (EVs) that have important roles in intercellular communication. They contain and carry bioactive molecules within their membranes which are delivered to target cells. Reproducible isolation and enrichment of these exosomes will aid in evaluation of cellular communication. We present an approach that involved the pre-processing of plasma, combined with ultracentrifugation (UC) and size exclusion chromatography (SEC) to isolate EVs and subsequently enrich exosomes. Four variations of this approach (denoted methods I to IV) were compared. Coupling an ultracentrifugation method with size exclusion chromatography (Method II) provided the best yield by nanoparticle tracking analyses (NTA), the presence of the exosomal markers CD63, Flotillin-1 and TSG-101 (immunoblotting) and showed exosome morphology using transmission electron microscopy (TEM). This method provides an efficient way to enrich the exosomes from blood (plasma), which could be potentially employed for clinical diagnostic assessment and therapeutic intervention.
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