Electrokinetically Driven Exosome Separation and Concentration Using Dielectrophoretic-Enhanced PDMS-Based Microfluidics

Ayala‐Mar, Sergio; Pérez-González, Víctor H.; Mata-Gómez, Marco A.; Gallo‐Villanueva, Roberto C.; González‐Valdez, José

doi:10.1021/acs.analchem.9b03448

Cited by 101 publications

(82 citation statements)

References 50 publications

(79 reference statements)

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“…Any voltage between 800 and 1000 V increased flow velocity and damaged the device. It was suggested that exosomes trapped by iDEP may be achievable at higher electric potentials [ 110 ]. Following this publication, Sergio et al [ 56 ] in 2019 enhanced the same device and applied 2000 V electric potential difference across the channel length.…”

Section: Discussionmentioning

confidence: 99%

Insulator Based Dielectrophoresis: Micro, Nano, and Molecular Scale Biological Applications

Benhal

Quashie

Kim

et al. 2020

Sensors

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Insulator based dielectrophoresis (iDEP) is becoming increasingly important in emerging biomolecular applications, including particle purification, fractionation, and separation. Compared to conventional electrode-based dielectrophoresis (eDEP) techniques, iDEP has been demonstrated to have a higher degree of selectivity of biological samples while also being less biologically intrusive. Over the past two decades, substantial technological advances have been made, enabling iDEP to be applied from micro, to nano and molecular scales. Soft particles, including cell organelles, viruses, proteins, and nucleic acids, have been manipulated using iDEP, enabling the exploration of subnanometer biological interactions. Recent investigations using this technique have demonstrated a wide range of applications, including biomarker screening, protein folding analysis, and molecular sensing. Here, we review current state-of-art research on iDEP systems and highlight potential future work.

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Section: Discussionmentioning

confidence: 99%

Insulator Based Dielectrophoresis: Micro, Nano, and Molecular Scale Biological Applications

Benhal

Quashie

Kim

et al. 2020

Sensors

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“…utilized a direct‐current insulator based dielectophoretic (DC‐iDEP) method to capture and separate exosomes of different sizes. [ 90 ] By using two electrically insulating post sections driven by iDEP, two subpopulations of exosomes can be successfully separated. For more classified exosome size sorting, Fujiwara et al.…”

Section: Microfluidics‐based Isolation Strategiesmentioning

confidence: 99%

Microfluidic‐Based Exosome Analysis for Liquid Biopsy

et al. 2021

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Liquid biopsy offers non‐invasive and real‐time molecular profiling of individual patients, and is thus considered a revolutionary technology in precision medicine. Exosomes have been acknowledged as significant biomarkers in liquid biopsy, as they play a central role in cell–cell communication and are closely related to the pathogenesis of most human malignancies. Nevertheless, in biofluids exosomes always co‐exist with other particles, and the cargo components of exosomes are highly heterogeneous. Thus, the isolation and molecular characterization of exosomes are still technically challenging. Microfluidics technology effectively addresses this challenge by virtue of its inherent advantages, such as precise manipulation of fluids, low consumption of samples and reagents, and a high level of integration. Recent advances in microfluidics allow in situ exosome capture and molecular detection with unprecedented selectivity and sensitivity. In this review, the state‐of‐the‐art developments in microfluidics‐based exosome research, including exosome isolation approaches and molecular detection strategies, with highlights of the characterization of exosomal biomarkers in cancer liquid biopsy is summarized. The major challenges are also discussed and some perspectives for the future directions of exosome‐based liquid biopsy in microfluidic systems are presented.

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“…[93] More recently, insulator-based DEP (iDEP), a variant of DEP, has gained attention because it does not require the insertion of electrodes in the chip, which can reduce costs and simplify the microfluidic device. [94,95] iDEP is the result of a non-uniform electric field between remote electrodes, implemented by embedding insulating posts in the microchannel. By adjusting the voltage magnitude, the DEP force exerted allows the separation of particles by sizes.…”

Section: Dielectrophoretic Isolation (Dep)mentioning

confidence: 99%

“…For instance, as shown in Figure 8, Ayala-Mar et al presented a direct current-iDEP (DC-iDEP) device with one channel and two electrically insulating posts for exosome isolation. [95] By adjusting the voltage applied across the main channel, larger (≈120 nm) and smaller (≈75 nm) exosomes can be trapped in the first and second parts, respectively. Exosome recovery is also possible in the side channels through electroosmotic flow.…”

Section: Dielectrophoretic Isolation (Dep)mentioning

confidence: 99%

A Holistic Review of the State‐of‐the‐Art Microfluidics for Exosome Separation: An Overview of the Current Status, Existing Obstacles, and Future Outlook

Ding

Yang

Gao

et al. 2021

Small

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Exosomes, which form a class of extracellular vesicles (EVs), are membrane-bound lipid nanovesicles with sizes typically in the Exosomes, a class of small extracellular vesicles (30-150 nm), are secreted by almost all types of cells into virtually all body fluids. These small vesicles are attracting increasing research attention owing to their potential for disease diagnosis and therapy. However, their inherent heterogeneity and the complexity of bio-fluids pose significant challenges for their isolation. Even the "gold standard," differential centrifugation, suffers from poor yields and is time-consuming. In this context, recent developments in microfluidic technologies have provided an ideal system for exosome extraction and these devices exhibit some fascinating properties such as high speeds, good portability, and low sample volumes. In this review, the focus is on the state-ofthe-art microfluidic technologies for exosome isolation and highlight potential directions for future research and development by analyzing the challenges faced by the current strategies. range of 30-150 nm. [1] They are secreted by almost all cells into diverse bio-fluids, including blood, urine, breast milk, saliva, lymph, and cerebrospinal fluid. [2] The discovery of exosomes dates back to the 1980s, but for many years after their discovery, they were regarded as "dust". [3,4] However, recent studies have shown that they play a crucial role in intercellular communication. [5,6] The biogenesis of exosomes includes double invagination of membranes, formation of intracellular multivesicular bodies (MVBs), and the release of exosomes (Figure 1). The first invagination process (plasma-membrane budding) generates early-sorting endosomes (ESEs) that can develop into late-sorting endosomes (LSEs). The LSEs are invaginated once again to form MVBs containing intraluminal vesicles (ILVs). Finally, the MVBs fuse with the plasma membrane to release ILVs (exosomes). [7] After release, these exosomes are taken up by recipient cells via multiple processes, such as macropinocytosis, fusion with the cell membrane, clathrin-dependent endocytosis, and phagocytosis. [8] The exosomes uptaken can act either at the surface of the recipient cells or deliver functional cargo in their bulk, thus affecting recipient-cell behavior. [9] Thus, exosomes play a key role in various physiological and pathological processes, including mammalian reproduction and development, immune responses, and disease progression. [10] Exosomes are reported to exhibit many excellent characteristics for clinical applications (Figure 2). 1) They can reflect the real-time state of the original cell, as they are actively secreted by living cells. 2) They are abundant in virtually all biological fluids (up to 10 10 vesicles per mL). [11] 3) Exosomes have enriched contents, including cell-surface substances and cytoplasmic constituents (including proteins, nucleic acids, lipids, and metabolites). Furthermore, exosomes can not only protect enzyme-sensitive cargos from degradation, but also ...

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Electrokinetically Driven Exosome Separation and Concentration Using Dielectrophoretic-Enhanced PDMS-Based Microfluidics

Cited by 101 publications

References 50 publications

Insulator Based Dielectrophoresis: Micro, Nano, and Molecular Scale Biological Applications

Insulator Based Dielectrophoresis: Micro, Nano, and Molecular Scale Biological Applications

Microfluidic‐Based Exosome Analysis for Liquid Biopsy

A Holistic Review of the State‐of‐the‐Art Microfluidics for Exosome Separation: An Overview of the Current Status, Existing Obstacles, and Future Outlook

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