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Purification and microRNA Profiling of Exosomes Derived from Blood and Culture Media
Published on: June 14, 2013
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MicroRNA Profiling of Exosomes
Melissa Daly1, Lorraine O'Driscoll2
1School of Pharmacy and Pharmaceutical Sciences & Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin 2, Ireland.
Methods in Molecular Biology (Clifton, N.J.)
|November 10, 2016
Summary
This study details methods for isolating and characterizing exosomes, which are nano-sized vesicles found in body fluids. These vesicles can transfer contents to other cells, making them promising circulating biomarkers.
Area of Science:
- Cell biology
- Biochemistry
- Biomarker discovery
Background:
- Exosomes are nano-sized vesicles secreted by various cells.
- They contain proteins and nucleic acids, transferable to recipient cells.
- Exosomes in body fluids are potential circulating biomarkers.
Purpose of the Study:
- To describe ultracentrifugation methods for exosome isolation from serum and plasma.
- To detail characterization techniques for isolated exosomes.
Main Methods:
- Ultracentrifugation for exosome isolation.
- Transmission electron microscopy (TEM) for morphology.
- Nanoparticle tracking analysis (NTA) for size and concentration.
- Immunoblotting for protein marker confirmation.
Main Results:
- Established ultracentrifugation protocols for serum and plasma exosome isolation.
- Confirmed exosome morphology, size, and protein markers using TEM, NTA, and immunoblotting.
Conclusions:
- Ultracentrifugation is a suitable method for exosome isolation from biofluids.
- Characterization techniques are essential for validating exosome purity and identity.
- Exosomes represent a valuable source for developing novel circulating biomarkers.
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