High-yield isolation of extracellular vesicles using aqueous two-phase system

Shin, Hyun–Woo; Han, Chul–Hi; Labuz, Joseph M.; Kim, Jiyoon; Kim, Jongmin; Cho, Siwoo; Gho, Yong Song; Takayama, Shuichi

doi:10.1038/srep13103

Cited by 120 publications

(94 citation statements)

References 39 publications

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“…MVs were isolated in accordance to protocol presented by Shin et al [21], which delivers high yield of extracellular vesicles using aqueous two-phase system composed of dextran (DEX) and polyethyleneglycol (PEG). Briefly, the culture medium is phase-separated by centrifugation at 1,000 ×g for 10 min at RT.…”

Section: Methodsmentioning

confidence: 99%

Macroautophagy and Selective Mitophagy Ameliorate Chondrogenic Differentiation Potential in Adipose Stem Cells of Equine Metabolic Syndrome: New Findings in the Field of Progenitor Cells Differentiation

Marycz

Kornicka

Grzesiak

et al. 2016

Oxidative Medicine and Cellular Longevity

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Equine metabolic syndrome (EMS) is mainly characterized by insulin resistance, obesity, and local or systemic inflammation. That unfriendly environment of adipose tissue has huge impact on stem cells population (ASC) residing within. In the present study, using molecular biology techniques and multiple imaging techniques (SEM, FIB-SEM, and confocal microscopy), we evaluated the impact of EMS on ASC viability and chondrogenic differentiation. Moreover, we visualized the mitochondrial network and dynamics in ASCCTRL and ASCEMS during control and chondrogenic conditions. In control conditions, ASCEMS were characterized by increased mitochondrial fission in comparison to ASCCTRL. We found that extensive remodeling of mitochondrial network including fusion and fission occurs during early step of differentiation. Moreover, we observed mitochondria morphology deterioration in ASCEMS. These conditions seem to cause autophagic shift in ASCEMS, as we observed increased accumulation of LAMP2 and formation of multiple autophagosomes in those cells, some of which contained dysfunctional mitochondria. “Autophagic” switch may be a rescue mechanism allowing ASCEMS to clear impaired by ROS proteins and mitochondria. Moreover it provides a precursors-to-macromolecules synthesis, especially during chondrogenesis. Our data indicates that autophagy in ASCEMS would be crucial for the quality control mechanisms and maintenance of cellular homeostasis ASCEMS allowing them to be in “stemness” status.

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Section: Methodsmentioning

confidence: 99%

Macroautophagy and Selective Mitophagy Ameliorate Chondrogenic Differentiation Potential in Adipose Stem Cells of Equine Metabolic Syndrome: New Findings in the Field of Progenitor Cells Differentiation

Marycz

Kornicka

Grzesiak

et al. 2016

Oxidative Medicine and Cellular Longevity

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“…Recently, in 2015, Shin et al, demonstrated an ATPS method for the purification of such extracellular vesicles. A polymer – polymer (PEG – dextran) ATPS was used which resulted in almost 70 % recovery just in a time span of 15 min [122]. …”

Section: Applications Of Aqueous Two-phase System (Atps)mentioning

confidence: 99%

Aqueous two-phase system (ATPS): an overview and advances in its applications

et al. 2016

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Aqueous two-phase system (ATPS) is a liquid-liquid fractionation technique and has gained an interest because of great potential for the extraction, separation, purification and enrichment of proteins, membranes, viruses, enzymes, nucleic acids and other biomolecules both in industry and academia. Although, the partition behavior involved in the method is complex and difficult to predict. Current research shows that it has also been successfully used in the detection of veterinary drug residues in food, separation of precious metals, sewage treatment and a variety of other purposes. The ATPS is able to give high recovery yield and is easily to scale up. It is also very economic and environment friendly method. The aim of this review is to overview the basics of ATPS, optimization and its applications.

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“…6) [82]. After samples are completely mixed with aqueous two-phase solution (polyethylene glycol (PEG) and dextran), the mixture is centrifuged at 3000×g for 10-30 min.…”

Section: Polymer Precipitationmentioning

confidence: 99%

“…ATPS is quick and easy, and does not requiring any incubation process [84]. In several biological fluids, ATPS shows higher recovery of EVs than does U/C and Exoquick ™ [82]. However, as EVs are isolated in a dextran-containing solution, so the effect of dextran must be reduced before ATPS can be widely accepted.…”

Section: Polymer Precipitationmentioning

confidence: 99%

Methods to isolate extracellular vesicles for diagnosis

Kang

Kim

Park

2017

Micro and Nano Syst Lett

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Extracellular vesicles (EVs) are small membrane-bound bodies that are released into extracellular space by diverse cells, and are found in body fluids like blood, urine and saliva. EVs contain RNA, DNA and proteins, which can be biomarkers for diagnosis. EVs can be obtained by minimally-invasive biopsy, so they are useful in disease diagnosis. High yield and purity contribute to precise diagnosis of disease, but damaged EVs and impurities can cause confused results. However, EV isolation methods have different yields and purities. Furthermore, the isolation method that is most suitable to maximize EV recovery efficiency depends on the experimental conditions. This review focuses on merits and demerits of several types of EV isolation methods, and provides examples of how to diagnose disease by exploiting information obtained by analysis of EVs.

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High-yield isolation of extracellular vesicles using aqueous two-phase system

Cited by 120 publications

References 39 publications

Macroautophagy and Selective Mitophagy Ameliorate Chondrogenic Differentiation Potential in Adipose Stem Cells of Equine Metabolic Syndrome: New Findings in the Field of Progenitor Cells Differentiation

Macroautophagy and Selective Mitophagy Ameliorate Chondrogenic Differentiation Potential in Adipose Stem Cells of Equine Metabolic Syndrome: New Findings in the Field of Progenitor Cells Differentiation

Aqueous two-phase system (ATPS): an overview and advances in its applications

Methods to isolate extracellular vesicles for diagnosis

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