A simplified method to recover urinary vesicles for clinical applications, and sample banking

Area of Science:

• Biochemistry
• Biotechnology
• Urology

Background:

• Urinary extracellular vesicles (EVs) are promising biomarkers for kidney and urogenital diseases.
• Current isolation methods, primarily ultracentrifugation, are laborious, time-consuming, and require large sample volumes, hindering widespread adoption.
• Challenges in EV isolation include protein contamination, vesicle loss, and storage issues, particularly for biobanking large cohorts.

Purpose of the Study:

• To develop a practical, cost-efficient, and scalable method for isolating urinary extracellular vesicles.
• To overcome the limitations of conventional ultracentrifugation protocols.
• To facilitate the utilization of urinary EVs in research and biobanking, especially in resource-limited settings.

Main Methods:

• Development of a novel vesicle isolation technique based on hydrostatic dialysis.
• Concentration of large urine volumes up to 1/100th of the original volume.
• Minimal interference from soluble proteins and reduced vesicle loss during isolation.

Main Results:

• The hydrostatic dialysis method yields easily manageable sample volumes.
• This technique is cost-efficient and eliminates the need for laborious and hazardous ultracentrifugation steps.
• Isolated vesicle fractions are suitable for various applications, including RNA analysis, outperforming conventional methods in yield.

Conclusions:

• Hydrostatic filtration dialysis offers a superior alternative for urinary EV isolation compared to ultracentrifugation.
• The method simplifies EV isolation, reducing labor, cost, and the need for specialized personnel and equipment.
• This technique is ideal for laboratories and biobanks working with urinary vesicles, promoting broader research and clinical applications.