Isolation of cancer-derived extracellular vesicle subpopulations by a size-selective microfluidic platform.

Zheyuan Chen,Yi Yang,Jun Kameoka,Hirohito Yamaguchi,Mien-Chi Hung

doi:10.1063/5.0008438

Abstract

Extracellular vesicles (EVs) play an important role in intercellular communication. Recently, there has been increasing interest in EVs as potential diagnostic biomarkers and therapeutic vehicles. However, the molecular properties and cargo information of EV subpopulations have not yet been fully investigated due to lack of reliable and reproducible EV separation technology. Current approaches have faced difficulties with efficiently isolating EVs from biofluids, especially subpopulations of small EVs. Here, we report an EV isolation method based on a size-selective microfluidic platform (ExoSMP) via nanomembrane filtration and electrophoretic force. This unique platform offers an enhanced approach to sorting a heterogeneous population of EVs based on size, with the additional advantages of being label-free and low-cost, and featuring a short processing time (<1 h), and convenient integration with downstream analysis. In this research, we used ExoSMP to demonstrate the isolation of cancer-derived small EVs (30–120 nm) with high recovery (94.2%) and reproducibility at an optimum sample flow rate. Furthermore, we investigated isolation of EV subpopulations by altering nanomembrane combinations with different pore size combinations (50 and 100 nm, 30 and 100 nm, 30 and 200 nm, and 30 and 50 nm). This ExoSMP technique can serve as a standardized EV isolation/separation tool, facilitating the clinical prospects of EVs and opening up a new avenue for future point-of-care applications in liquid biopsies.

Highlights

Among translational and clinical researchers, there is significant interest in extracellular vesicles (EVs) as potential diagnostic biomarkers and therapeutic vehicles that can be fast-tracked to clinical evaluation and precision medicine applications.[1,2] Extracellular vesicles (EVs) refer to a heterogenous lipid bilayer of membranous structures that hold information in the form of proteins, lipids, or nucleic acids, thereby physiologically and pathologically influencing the intercellular communication of both the recipient and parent cells.[3]
This unique platform offers an enhanced approach to sorting a heterogeneous population of EVs based on size, with the additional advantages of being label-free and low-cost, and featuring a short processing time (
We developed and demonstrated a size-selective microfluidic platform (ExoSMP) for automated, consistent, and reliable EV isolation

Summary

Introduction

Among translational and clinical researchers, there is significant interest in extracellular vesicles (EVs) as potential diagnostic biomarkers and therapeutic vehicles that can be fast-tracked to clinical evaluation and precision medicine applications.[1,2] EVs refer to a heterogenous lipid bilayer of membranous structures that hold information in the form of proteins, lipids, or nucleic acids, thereby physiologically and pathologically influencing the intercellular communication of both the recipient and parent cells.[3] EVs are released by most viable cells These particles can be isolated and collected from various bodily fluids such as blood, saliva, urine, lymph, and milk. In the past few decades, there are at least three main subgroups that have been identified and termed as exosomes, microvesicles, and apoptotic bodies.[4,5] The existing criteria adopted to discriminate among the subpopulations are based on size, density, function, and molecular cargo.[6] The physical properties of EVs

Methods

Results

Conclusion