Microfluidic Approaches and Methods Enabling Extracellular Vesicle Isolation for Cancer Diagnostics

Singh, Premanshu Kumar; Patel, Aavishkar A.; Kaffenes, Anastasia; Hord, Catherine; Kesterson, Delaney; Prakash, Shaurya

doi:10.3390/mi13010139

Cited by 12 publications

(8 citation statements)

References 141 publications

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“…It was di cult to detect enough metabolites to make a conclusion for exosomal metabolites. It is also important to discuss the fact that no single standardized method exists yet for the perfect isolation of exosomes for their use in cancer biomarker studies (94)(95)(96). There is still some debate as to whether it is even possible to isolate the intact and pure exosomes.…”

Section: Discussionmentioning

confidence: 99%

Source of Liquid Biopsy Biomarker: Exosome vs Whole Plasma, Fasting vs Non-fasting

Nasu

Khadka

Jijiwa

et al. 2022

Preprint

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Background The liquid biopsy using plasma samples is being studied to find biomarkers for clinical applications. Exosomes encompass nucleic acids and metabolites that have been highlighted as a potential biomarker source. To test the efficacy of exosomes over plasma, we compared the profiles of small non-coding RNAs (ncRNAs) and metabolites extracted from exosomes (which were purified from plasma) to the profiles extracted directly from whole plasma. The fasting and non-fasting status of the samples were also compared. Results We found that ncRNA profiles were not affected by fasting for both exosomal and plasma samples. Our results showed that ncRNAs extracted from exosomes were found to have the more consistent profiles between fasting and non-fasting samples. The whole plasma RNA profiles contained high concentrations of cell-derived miRNAs that were likely based on hemolysis. We also found that some metabolites in whole plasma showed significant changes in concentration due to fasting status, whereas others did not. Conclusions Here, we propose that 1) fasting isn’t necessary for liquid biopsy study for both circulating ncRNA and metabolomic profiling as long as metabolites which aren’t affected by fasting status are chosen. 2) Exosomal RNAs must be used to obtain consistent results without batch effects in plasma samples due to different levels of hemolysis.

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Section: Discussionmentioning

confidence: 99%

Source of Liquid Biopsy Biomarker: Exosome vs Whole Plasma, Fasting vs Non-fasting

Nasu

Khadka

Jijiwa

et al. 2022

Preprint

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“…Microfluidics not only have the advantage of reducing the consumption of samples and reagents and providing higher resolution and sensitivity, but they also significantly reduce the analysis time and greatly improve the analysis efficiency [ 42 , 43 ]. Due to the vast amounts of progress in microfabrication technology through recent years, microfluidic technology has become a promising method for exosome isolation [ 44 ]. Currently, microfluidic-based exosome-separation technologies can be classified into two types: (I) physical property-based microfluidic for exosome separation, (II) immunoaffinity-based microfluidic for exosome separation.…”

Section: Microfluidic-based Technology For Exosome Isolationmentioning

confidence: 99%

Microfluidic Technology for the Isolation and Analysis of Exosomes

Wang

et al. 2022

Micromachines

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Exosomes are lipid-bilayer enclosed vesicles with diameters of 30–150 nm, which play a pivotal role in cell communication by transporting their cargoes such as proteins, lipids, and genetic materials. In recent years, exosomes have been under intense investigation, as they show great promise in numerous areas, especially as bio-markers in liquid biopsies. However, due to the high heterogeneity and the nano size of exosomes, the separation of exosomes is not easy. This review will deliver an outline of the conventional methods and the microfluidic-based technologies for exosome separation. Particular attention is devoted to microfluidic devices, highlighting the efficiency of exosome isolation by these methods. Additionally, this review will introduce advances made in the integrated microfluidics technologies that enable the separation and analysis of exosomes.

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“…They have been recently evaluated on human plasma and CSF by Ter-Ovanesyan et al [ 6 ]. Moreover, new methods are emerging, e.g., microfluidic devices [ 7 , 8 ], clog-free ultrafiltration (EXODUS) [ 9 ] and nanomaterial-based isolations [ 10 ]. Each method has its advantages and disadvantages and needs to be optimized according to the biofluid and the research hypothesis.…”

Section: Introductionmentioning

confidence: 99%

Unveiling the Native Morphology of Extracellular Vesicles from Human Cerebrospinal Fluid by Atomic Force and Cryogenic Electron Microscopy

et al. 2022

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Extracellular vesicles (EVs) are membranous structures in biofluids with enormous diagnostic/prognostic potential for application in liquid biopsies. Any such downstream application requires a detailed characterization of EV concentration, size and morphology. This study aimed to observe the native morphology of EVs in human cerebrospinal fluid after traumatic brain injury. Therefore, they were separated by gravity-driven size-exclusion chromatography (SEC) and investigated by atomic force microscopy (AFM) in liquid and cryogenic transmission electron microscopy (cryo-TEM). The enrichment of EVs in early SEC fractions was confirmed by immunoblot for transmembrane proteins CD9 and CD81. These fractions were then pooled, and the concentration and particle size distribution were determined by Tunable Resistive Pulse Sensing (around 1010 particles/mL, mode 100 nm) and Nanoparticle Tracking Analysis (around 109 particles/mL, mode 150 nm). Liquid AFM and cryo-TEM investigations showed mode sizes of about 60 and 90 nm, respectively, and various morphology features. AFM revealed round, concave, multilobed EV structures; and cryo-TEM identified single, double and multi-membrane EVs. By combining AFM for the surface morphology investigation and cryo-TEM for internal structure differentiation, EV morphological subpopulations in cerebrospinal fluid could be identified. These subpopulations should be further investigated because they could have different biological functions.

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Microfluidic Approaches and Methods Enabling Extracellular Vesicle Isolation for Cancer Diagnostics

Cited by 12 publications

References 141 publications

Source of Liquid Biopsy Biomarker: Exosome vs Whole Plasma, Fasting vs Non-fasting

Source of Liquid Biopsy Biomarker: Exosome vs Whole Plasma, Fasting vs Non-fasting

Microfluidic Technology for the Isolation and Analysis of Exosomes

Unveiling the Native Morphology of Extracellular Vesicles from Human Cerebrospinal Fluid by Atomic Force and Cryogenic Electron Microscopy

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