Comparison of isolation methods of exosomes and exosomal RNA from cell culture medium and serum

Tang, Yueting; Huang, Yiyao; Zheng, Lei; Qin, Sihua; Xu, Xuping; An, Tongqing; Xu, Yong; Wu, Yangyang; Hu, Xiumei; Ping, Baohong; Wang, Qian

doi:10.3892/ijmm.2017.3080

Cited by 375 publications

(294 citation statements)

References 27 publications

Supporting

Mentioning

273

Contrasting

Unclassified

Order By: Relevance

“…Several previous publications reported the feasibility and utility of sequencing small RNA in EVs isolated from serum, plasma, urine, and cell culture supernatant [2,27–30]. Small RNA-Seq experiments often focus on valuable applications such as liquid biopsy-based diagnostics and, consequently, clinical samples.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of serum extracellular vesicle isolation methods for profiling miRNAs by next‐generation sequencing

Buschmann

Kirchner

Hermann

et al. 2018

J of Extracellular Vesicle

203

165

View full text Add to dashboard Cite

Extracellular vesicles (EVs) are intercellular communicators with key functions in physiological and pathological processes and have recently garnered interest because of their diagnostic and therapeutic potential. The past decade has brought about the development and commercialization of a wide array of methods to isolate EVs from serum. Which subpopulations of EVs are captured strongly depends on the isolation method, which in turn determines how suitable resulting samples are for various downstream applications. To help clinicians and scientists choose the most appropriate approach for their experiments, isolation methods need to be comparatively characterized. Few attempts have been made to comprehensively analyse vesicular microRNAs (miRNAs) in patient biofluids for biomarker studies. To address this discrepancy, we set out to benchmark the performance of several isolation principles for serum EVs in healthy individuals and critically ill patients. Here, we compared five different methods of EV isolation in combination with two RNA extraction methods regarding their suitability for biomarker discovery-focused miRNA sequencing as well as biological characteristics of captured vesicles. Our findings reveal striking method-specific differences in both the properties of isolated vesicles and the ability of associated miRNAs to serve in biomarker research. While isolation by precipitation and membrane affinity was highly suitable for miRNA-based biomarker discovery, methods based on size-exclusion chromatography failed to separate patients from healthy volunteers. Isolated vesicles differed in size, quantity, purity and composition, indicating that each method captured distinctive populations of EVs as well as additional contaminants. Even though the focus of this work was on transcriptomic profiling of EV-miRNAs, our insights also apply to additional areas of research. We provide guidance for navigating the multitude of EV isolation methods available today and help researchers and clinicians make an informed choice about which strategy to use for experiments involving critically ill patients.

show abstract

Section: Introductionmentioning

confidence: 99%

Evaluation of serum extracellular vesicle isolation methods for profiling miRNAs by next‐generation sequencing

Buschmann

Kirchner

Hermann

et al. 2018

J of Extracellular Vesicle

203

165

View full text Add to dashboard Cite

show abstract

“…The conditioned medium was ultra‐centrifuged at 100,000 g for 70 minutes at 4°C as described previously . The exosomes pellets were resuspended in PBS and stored at −80°C.…”

Section: Methodsmentioning

confidence: 99%

Human mesenchymal‐stem‐cells‐derived exosomes are important in enhancing porcine islet resistance to hypoxia

Nie

Yang

et al. 2018

Xenotransplantation

View full text Add to dashboard Cite

show abstract

“…Airway EVs from BALF of mice were isolated using the ExoQuick Exosome Precipitation Solution. Compared to the usual technique of ultracentrifugation, this provides a higher extraction efficiency but has the risk of protein contamination when used to isolate exosomes in cell culture media . The results showed that the amount of EVs observed increased by 8.9‐fold in the BALF of HDM‐exposed mice compared to the control group.…”

Section: Resultsmentioning

confidence: 99%

“…Compared to the usual technique of ultracentrifugation, this provides a higher extraction efficiency but has the risk of protein contamination when used to isolate exosomes in cell culture media. 82 The results showed that the amount of EVs observed increased by…”

Section: Gon Et Al 81 Analysed Variations In the Production Of Airwaymentioning

confidence: 97%

Extracellular vesicles and asthma: A review of the literature

Sangaphunchai

Todd

Fairclough

2020

Clin Experimental Allergy

View full text Add to dashboard Cite

Asthma is a chronic, recurrent and incurable allergy‐related respiratory disease characterized by inflammation, bronchial hyperresponsiveness and narrowing of the airways. Extracellular vesicles (EVs) are a universal feature of cellular function and can be detected in different bodily fluids. Recent evidence has shown the possibility of using EVs in understanding the pathogenesis of asthma, including their potential as diagnostic and therapeutic tools. Studies have reported that EVs released from key cells involved in asthma can induce priming and activation of other asthma‐associated cells. A literature review was conducted on all current research regarding the role and function of EVs in the pathogenesis of asthma via the PRISMA statement method. An electronic search was performed using EMBASE and PubMed through to November 2018. The EMBASE search returned 76 papers, while the PubMed search returned 211 papers. Following duplicate removal, titles and abstracts were screened for eligibility with a total of 34 studies included in the final qualitative analysis. The review found evidence of association between the presence of EVs and physiological changes characteristic of asthma, suggesting that EVs are involved in the pathogenesis, with the weight of evidence presently favouring deleterious effects of EVs in asthma. Numerous studies highlighted differences in exosomal contents between EVs of healthy and asthmatic individuals, which could be employed as potential diagnostic markers. In some circumstances, EVs were also found to be suppressive to disease, but more often promote inflammation and airway remodelling. In conclusion, EVs hold immense potential in understanding the pathophysiology of asthma, and as diagnostic and therapeutic markers. While more research is needed for definitive conclusions and their application in medical practice, the literature presented in this review should encourage further research and discovery within the field of EVs and asthma.

show abstract

Comparison of isolation methods of exosomes and exosomal RNA from cell culture medium and serum

Cited by 375 publications

References 27 publications

Evaluation of serum extracellular vesicle isolation methods for profiling miRNAs by next‐generation sequencing

Evaluation of serum extracellular vesicle isolation methods for profiling miRNAs by next‐generation sequencing

Human mesenchymal‐stem‐cells‐derived exosomes are important in enhancing porcine islet resistance to hypoxia

Extracellular vesicles and asthma: A review of the literature

Contact Info

Product

Resources

About