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Updated: Jul 21, 2025
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Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis
Published on: January 7, 2019
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Optimized procedure for high-throughput transcriptome profiling of small extracellular vesicles isolated from low
Petra Vychytilova-Faltejskova1, Sara Vilmanova1, Lucie Pifkova1
1Centre for Molecular Medicine, Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
Clinical Chemistry and Laboratory Medicine
|July 28, 2023
Summary
Optimized protocols for isolating small extracellular vesicles (EVs) and their RNA from serum enable high-throughput transcriptome analysis. This approach identified numerous deregulated non-coding RNAs in colorectal cancer patients, highlighting their potential as biomarkers.
Area of Science:
- Biochemistry
- Molecular Biology
- Genomics
Background:
- Small extracellular vesicles (EVs) are key mediators of intercellular communication and hold promise as biomarker sources.
- Standardization of small EV analysis is lacking, hindering clinical translation.
Purpose of the Study:
- To compare different high-throughput methods for analyzing the transcriptome of small EVs.
- To optimize protocols for small EV RNA isolation from low serum volumes.
Main Methods:
- Small EVs were isolated from serum using size exclusion chromatography.
- RNA extraction efficiency was compared using different kits, with Monarch Total RNA Miniprep Kit showing optimal results.
- Whole transcriptome analysis was performed using microarrays and RT-qPCR.
Main Results:
- Size exclusion chromatography proved suitable for small EV isolation.
- 7,182 deregulated transcripts were identified between colorectal cancer patients and healthy controls.
- Non-coding RNAs, particularly lncRNAs, constituted the majority of deregulated transcripts.
Conclusions:
- An optimized protocol for small EV and RNA isolation from low serum volumes was developed.
- Clariom D Pico Assays are suitable for transcriptome profiling of small EVs.
- The findings support the potential of small EV non-coding RNAs as biomarkers for colorectal cancer.
