Single Step Isolation of Extracellular Vesicles from Large-Volume Samples with a Bifurcated A4F Microfluidic Device

Miks Priedols; Gunita Paidere; Pauls Kaukis; Cristina Bajo-Santos; Arnita Spule; Antons Miscenko; Gatis Mozolevskis; Roberts Rimsa; Arturs Abols

doi:10.3791/66019

Single Step Isolation of Extracellular Vesicles from Large-Volume Samples with a Bifurcated A4F M...

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Bioengineering

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JoVE Journal Bioengineering

Single Step Isolation of Extracellular Vesicles from Large-Volume Samples with a Bifurcated A4F Microfluidic Device

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06:28 min

February 2, 2024

DOI: 10.3791/66019-v

Miks Priedols*¹, Gunita Paidere*², Pauls Kaukis*¹, Cristina Bajo-Santos¹, Arnita Spule^3,4, Antons Miscenko¹, Gatis Mozolevskis^2,3, Roberts Rimsa^2,3, Arturs Abols^1,3

¹Latvian Biomedical Research and Study Centre, ²Institute of Solid-State Physics,University of Latvia, ³Cellbox labs LTD, Atari, ⁴Faculty of Materials Science and Applied Chemistry, Institute of General Chemical Engineering,Riga Technical university

Overview

This study presents a microfluidic device designed for the direct isolation of extracellular vesicles from large volumes of biofluids. The device allows for continuous flow and automation, addressing the challenges of current isolation methods.

Key Study Components

Area of Science

Biomedical applications
Extracellular vesicle isolation
Microfluidics

Background

Extracellular vesicles have significant potential in various biomedical fields.
Current isolation methods are often impractical for clinical use.
Isolating vesicles from large biofluid samples is particularly challenging.
Existing methods require prior collection and processing of biofluids.

Purpose of Study

To develop a microfluidic device for efficient isolation of extracellular vesicles.
To enable integration with bioreactors for continuous processing.
To reduce the labor and time involved in current isolation techniques.

Methods Used

Asymmetric field-flow fractionation for vesicle isolation.
Design of a microfluidic device for continuous flow.
Testing the impact of liquid viscosity on isolation efficiency.
Integration of the device with bioreactor systems.

Main Results

The device successfully isolates extracellular vesicles from large-volume biofluids.
Continuous flow operation was achieved, facilitating automation.
Liquid viscosity was found to influence the isolation process.
The method reduces the need for prior biofluid processing.

Conclusions

The microfluidic device presents a viable solution for extracellular vesicle isolation.
It enhances the practicality of using vesicles in clinical applications.
Future work may focus on optimizing the device for various biofluids.

Frequently Asked Questions

What are extracellular vesicles?

Extracellular vesicles are small membrane-bound particles released by cells that play a role in cell communication and can carry biomolecules.

How does the microfluidic device work?

The device uses asymmetric field-flow fractionation to isolate extracellular vesicles from biofluids in a continuous flow manner.

What is the significance of isolating extracellular vesicles?

Isolating extracellular vesicles is crucial for their use in diagnostics and therapeutics in various biomedical applications.

What challenges do current isolation methods face?

Current methods are often time-consuming, labor-intensive, and not suitable for large-volume samples.

How does viscosity affect vesicle isolation?

Liquid viscosity can impact the efficiency of the isolation process, influencing the yield and purity of the vesicles obtained.

Extracellular vesicles hold immense promise for biomedical applications, but current isolation methods are time-consuming and impractical for clinical use. In this study, we present a microfluidic device that enables the direct isolation of extracellular vesicles from large volumes of biofluids in a continuous manner with minimal steps.

We see the potential of extracellular vesicles, but also recognize the difficulty in their isolation, especially from large sample of biofluids such as bioreactor medium. In this research we tested if we can use asymmetric field-flow field fractionation to isolate the extracellular vesicles. As of now, isolating extracellular vesicles directly from biofluids in a continuous flow for integration into bioreactors is impossible.

Instead, biofluids must be first collected and processed, adding viability and labor. Our device, designed for extracellular vesicle isolation from large-volume biofluids, enables integration with bioreactors for continuous flow and automation. We found liquid viscosity might impact extracellular vesicles isolation using asymmetric field-flow fractionation.

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