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Microfluidic device (ExoChip) for on-chip isolation, quantification and characterization of circulating exosomes
Shailender Singh Kanwar1, Christopher James Dunlay, Diane M Simeone
1Department of Chemical Engineering, College of Engineering University of Michigan, 2300 Hayward Street, Ann Arbor, Michigan-48109, USA. snagrath@umich.edu.
Lab on a Chip
|April 12, 2014
Summary
A novel microfluidic device, ExoChip, rapidly isolates and quantifies circulating exosomes from blood serum. This platform shows significantly higher exosome capture in pancreatic cancer patients, offering a promising tool for cancer biomarker discovery.
Area of Science:
- Biotechnology
- Nanotechnology
- Cancer Research
Background:
- Extracellular vesicles, including exosomes, are released by cells and circulate in bodily fluids.
- Circulating exosomes are potential biomarkers for disease detection.
- Current methods for exosome isolation and analysis are often slow and not high-throughput.
Purpose of the Study:
- To develop a rapid, low-cost, and reproducible microfluidic platform for isolating and quantifying circulating exosomes (cirEVs).
- To assess the platform's efficacy in distinguishing between exosomes from healthy individuals and pancreatic cancer patients.
Main Methods:
- Fabrication of a microfluidic device (ExoChip) using polydimethylsiloxane (PDMS).
- Functionalization of ExoChip with antibodies against CD63 for specific exosome capture.
- Staining captured exosomes with a fluorescent dye (DiO) for quantification via plate-reader.
- Confirmation of exosome identity using immuno-electron-microscopy and Western blotting.
- Analysis of exosomal RNA integrity for microRNA profiling.
Main Results:
- ExoChip successfully isolated and quantified circulating exosomes from human blood serum.
- A statistically significant 2.34-fold increase in captured exosomes was observed in pancreatic cancer patients compared to healthy individuals.
- Immuno-electron-microscopy and Western blotting confirmed the exosomal origin of captured vesicles.
- Intact RNA was recovered from exosomes, enabling microRNA profiling.
Conclusions:
- ExoChip is a simple, cost-effective, and high-throughput platform for exosome isolation and quantification.
- The platform demonstrates potential for early cancer detection through exosome biomarker analysis.
- ExoChip is suitable for molecular screening in cancer diagnostics and research.
