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Related Concept Videos

Overview of Exosomes01:36

Overview of Exosomes

Exosomes are stable, lipid bilayer-enclosed vesicles capable of crossing biological barriers. They can carry a wide range of molecules required for intercellular communication. Once exosomes are released from the cell where they originated, they enter a recipient cell through various pathways such as fusion, receptor-mediated endocytosis, macropinocytosis, and phagocytosis.
Stahl et al. discovered exosomes in 1983, but the exosomes were initially considered waste products released from the...
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Isolation and Characterization of RNA-Containing Exosomes
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Comparing Two Methods for the Isolation of Exosomes.

Mohammad A Aziz1, Benedict Seo1, Haizal M Hussaini1

  • 1Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, PO Box 56, Dunedin 9054, New Zealand.

Journal of Nucleic Acids
|November 4, 2022
PubMed
Summary

Ultracentrifugation (UC) yields higher RNA yields from exosomes compared to the Exoquick-TC PLUS™ kit. UC is preferred for mRNA analysis, though the kit is a viable alternative when UC is unavailable.

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Area of Science:

  • Extracellular vesicles research
  • Nanotechnology in diagnostics
  • Molecular biology techniques

Background:

  • Exosomes are key nanovesicles involved in intercellular communication.
  • Exosomal RNA, particularly mRNA, holds diagnostic potential for various diseases.
  • Efficient exosome isolation is crucial for downstream molecular analysis.

Purpose of the Study:

  • To compare the efficacy of two exosome isolation methods: Exoquick-TC PLUS™ kit vs. ultracentrifugation (UC).
  • To evaluate the impact of isolation method on RNA yield and quality for diagnostic applications.

Main Methods:

  • Exosome isolation using Exoquick-TC PLUS™ kit and ultracentrifugation (UC).
  • Characterization via cryo-electron microscopy and dynamic light scattering (DLS).
  • Quantification of total RNA and mRNA using qPCR.

Main Results:

  • Both methods yielded exosomes (50-150 nm diameter).
  • UC exosomes showed a single peak in DLS; kit exosomes showed multiple peaks, suggesting impurities.
  • UC exosomes contained significantly higher total RNA and mRNA (lower Ct values) compared to kit exosomes (P < 0.0001).

Conclusions:

  • Ultracentrifugation (UC) is superior for exosome isolation when high RNA yield is critical for mRNA analysis.
  • The Exoquick-TC PLUS™ kit is a practical alternative, successfully quantifying four out of five selected genes, especially when UC is not feasible.