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Isolation and Profiling of MicroRNA-containing Exosomes from Human Bile
Published on: June 13, 2016
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Low pH increases the yield of exosome isolation
Jae-Jun Ban1, Mijung Lee2, Wooseok Im2
1Department of Bioscience and Biotechnology, College of Life Science, Institute of Biotechnology, Sejong University, Seoul, South Korea.
Biochemical and Biophysical Research Communications
|April 8, 2015
Summary
Acidic conditions enhance the yield and stability of exosomes, crucial vesicles for intercellular communication. This study reveals that low pH favors exosome existence and isolation, impacting cancer research.
Area of Science:
- Extracellular Vesicles Research
- Cancer Biology
- Biochemistry
Background:
- Exosomes are key mediators of intercellular communication, delivering molecules between cells.
- Cancer-derived exosomes are implicated in tumor progression and metastasis.
- The tumor microenvironment's acidic pH influences exosome function, yet its impact on exosome isolation is understudied.
Purpose of the Study:
- To investigate the effect of pH on exosome stability and isolation yield.
- To determine optimal pH conditions for exosome protein and RNA recovery.
- To analyze the expression of exosome markers under varying pH conditions.
Main Methods:
- Incubation of exosomes in acidic, neutral, and alkaline conditioned media.
- Quantification of total exosomal protein and RNA.
- Western blot analysis of exosome markers (CD9, CD63, HSP70).
Main Results:
- Exosomal protein and nucleic acid concentrations significantly increased in acidic conditions compared to neutral.
- Elevated levels of exosome markers (CD9, CD63, HSP70) were observed in acidic environments.
- No exosomal protein, RNA, or markers were detected in alkaline conditions.
Conclusions:
- Acidic conditions are favorable for the existence and isolation of exosomes.
- pH significantly impacts exosome yield and marker detection.
- This finding is crucial for standardizing exosome isolation protocols in research.
