Extracellular Vesicles: Isolation Methods

Pariset, Eloïse; Agache, Vincent; Millet, Arnaud

doi:10.1002/adbi.201700040

Cited by 30 publications

(18 citation statements)

References 86 publications

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“…Ultracentrifugation (UC) is currently the gold-standard in exosome isolation and widely used in laboratories, 136 where roughly 56% of all exosome isolation is performed using this technique 150 . UC is based on the difference in the sedimentation rate of particles, which is affected by their size, density, and shape 151 . By applying centrifugal forces, sample components can be separated sequentially according to their physical properties, and the density and viscosity of the solvent.…”

Section: Isolationmentioning

confidence: 99%

Challenges and opportunities in exosome research—Perspectives from biology, engineering, and cancer therapy

et al. 2019

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Exosomes are small (∼30–140 nm) lipid bilayer-enclosed particles of endosomal origin. They are a subset of extracellular vesicles (EVs) that are secreted by most cell types. There has been growing interest in exosome research in the last decade due to their emerging role as intercellular messengers and their potential in disease diagnosis. Indeed, exosomes contain proteins, lipids, and RNAs that are specific to their cell origin and could deliver cargo to both nearby and distant cells. As a result, investigation of exosome cargo contents could offer opportunities for disease detection and treatment. Moreover, exosomes have been explored as natural drug delivery vehicles since they can travel safely in extracellular fluids and deliver cargo to destined cells with high specificity and efficiency. Despite significant efforts made in this relatively new field of research, progress has been held back by challenges such as inefficient separation methods, difficulties in characterization, and lack of specific biomarkers. In this review, we summarize the current knowledge in exosome biogenesis, their roles in disease progression, and therapeutic applications and opportunities in bioengineering. Furthermore, we highlight the established and emerging technological developments in exosome isolation and characterization. We aim to consider critical challenges in exosome research and provide directions for future studies.

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Section: Isolationmentioning

confidence: 99%

Challenges and opportunities in exosome research—Perspectives from biology, engineering, and cancer therapy

et al. 2019

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“…Since its first introduction in 2004 by Huang et al . 1 , Deterministic Lateral Displacement (DLD) has been extensively implemented and improved 2–4 to purify biological components, while preserving their integrity, such as blood cells 5–8 , circulating tumor cells 9–11 , bacteria 12 , parasites 13 , spores 14 and even nanoparticles 15–17 . In DLD devices, passive and label-free separation is induced by an array of regularly arranged pillars that give specific trajectories to particles of different sizes.…”

Section: Introductionmentioning

confidence: 99%

Separation of Biological Particles in a Modular Platform of Cascaded Deterministic Lateral Displacement Modules

Pariset

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Fouillet

et al. 2018

Sci Rep

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Deterministic lateral displacement (DLD) has been extensively implemented in the last decade for size-based sample preparation, owing to its high separation performances for a wide range of particle dimensions. However, separating particles from 1 μm to 10 μm in one single DLD device is challenging because of the required diversity of pillar dimensions and inherent fabrication issues. This paper presents an alternative approach to achieve the extraction of E. coli bacteria from blood samples spiked with prostate cancer cells. Our approach consists in cascading individual DLD devices in a single automated platform, using flexible chambers that successively collect and inject the sample between each DLD stage without any external sample manipulation. Operating DLD separations independently enables to maximize the sorting efficiency at each step, without any disturbance from downstream stages. The proposed two-step automated protocol is applied to the separation of three types of components (bacteria, blood particles and cancer cells), with a depletion yield of 100% for cancer cells and 93% for red blood cells. This cascaded approach is presented for the first time with two DLD modules and is upscalable to improve the dynamic range of currently available DLD devices.

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“…As a passive and label-free purification technique, DLD has a high potential to address both micrometer- and nanometer-sized biological particles. Examples of applications are the isolation of circulating tumor cells (CTCs) from blood samples [ 1 ][ 2 ][ 3 ][ 4 ][ 5 ], the extraction of white blood cells (WBCs) [ 6 ][ 7 ][ 8 ][ 9 ][ 10 ][ 11 ], the manipulation of red blood cells (RBCs) [ 12 ][ 13 ][ 14 ][ 15 ][ 16 ][ 17 ], the isolation of cells [ 18 ][ 19 ], platelets [ 20 ], fungal spores [ 21 ], bacteria [ 22 ], parasites [ 23 ], and the fractionation of isolated extracellular vesicles into several subpopulations [ 24 ][ 25 ].…”

Section: Introductionmentioning

confidence: 99%

Purification of complex samples: Implementation of a modular and reconfigurable droplet-based microfluidic platform with cascaded deterministic lateral displacement separation modules

et al. 2018

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Particle separation in microfluidic devices is a common problematic for sample preparation in biology. Deterministic lateral displacement (DLD) is efficiently implemented as a size-based fractionation technique to separate two populations of particles around a specific size. However, real biological samples contain components of many different sizes and a single DLD separation step is not sufficient to purify these complex samples. When connecting several DLD modules in series, pressure balancing at the DLD outlets of each step becomes critical to ensure an optimal separation efficiency. A generic microfluidic platform is presented in this paper to optimize pressure balancing, when DLD separation is connected either to another DLD module or to a different microfluidic function. This is made possible by generating droplets at T-junctions connected to the DLD outlets. Droplets act as pressure controllers, which perform at the same time the encapsulation of DLD sorted particles and the balance of output pressures. The optimized pressures to apply on DLD modules and on T-junctions are determined by a general model that ensures the equilibrium of the entire platform. The proposed separation platform is completely modular and reconfigurable since the same predictive model applies to any cascaded DLD modules of the droplet-based cartridge.

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Extracellular Vesicles: Isolation Methods

Cited by 30 publications

References 86 publications

Challenges and opportunities in exosome research—Perspectives from biology, engineering, and cancer therapy

Challenges and opportunities in exosome research—Perspectives from biology, engineering, and cancer therapy

Separation of Biological Particles in a Modular Platform of Cascaded Deterministic Lateral Displacement Modules

Purification of complex samples: Implementation of a modular and reconfigurable droplet-based microfluidic platform with cascaded deterministic lateral displacement separation modules

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