Improved isolation strategies to increase the yield and purity of human urinary exosomes for biomarker discovery.

Hashemi Gheinani, Ali; Vögeli, Mike; Baumgartner, Ulrich; Vassella, Erik; Draeger, Annette; Burkhard, Fiona C.; Monastyrskaya-Stäuber, Katia (2018). Improved isolation strategies to increase the yield and purity of human urinary exosomes for biomarker discovery. Scientific Reports, 8(1), p. 3945. Nature Publishing Group 10.1038/s41598-018-22142-x

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Circulating miRNAs are detected in extracellular space and body fluids such as urine. Circulating RNAs can be packaged in secreted urinary extracellular vesicles (uEVs) and thus protected from degradation. Urinary exosome preparations might contain specific miRNAs, relevant as biomarkers in renal and bladder diseases. Major difficulties in application of uEVs into the clinical environment are the high variability and low reproducibility of uEV isolation methods. Here we used five different methods to isolate uEVs and compared the size distribution, morphology, yield, presence of exosomal protein markers and RNA content of uEVs. We present an optimized ultracentrifugation and size exclusion chromatography approach for highly reproducible isolation for 50-150 nm uEVs, corresponding to the exosomes, from 50 ml urine. We profiled the miRNA content of uEVs and total urine from the same samples with the NanoString platform and validated the data using qPCR. Our results indicate that 18 miRNAs, robustly detected in uEVs were always present in the total urine. However, 15 miRNAs could be detected only in the total urine preparations and might represent naked circulating miRNA species. This is a novel unbiased and reproducible strategy for uEVs isolation, content normalization and miRNA cargo analysis, suitable for biomarker discovery studies.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Anatomy
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > DBMR Forschung Mu35 > Forschungsgruppe Urologie
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > DBMR Forschung Mu35 > Forschungsgruppe Urologie

04 Faculty of Medicine > Service Sector > Institute of Pathology
04 Faculty of Medicine > Department of Dermatology, Urology, Rheumatology, Nephrology, Osteoporosis (DURN) > Clinic of Urology
04 Faculty of Medicine > Service Sector > Institute of Pathology > Tumour Pathology
09 Interdisciplinary Units > Microscopy Imaging Center MIC

UniBE Contributor:

Hashemi Gheinani, Ali; Baumgartner, Ulrich; Vassella, Erik; Draeger, Annette; Burkhard, Fiona C. and Monastyrskaya-Stäuber, Katia

Subjects:

600 Technology > 610 Medicine & health
500 Science > 570 Life sciences; biology

ISSN:

2045-2322

Publisher:

Nature Publishing Group

Language:

English

Submitter:

Laetitia Hayoz

Date Deposited:

05 Jul 2018 10:44

Last Modified:

12 Jul 2019 16:06

Publisher DOI:

10.1038/s41598-018-22142-x

PubMed ID:

29500443

BORIS DOI:

10.7892/boris.118239

URI:

https://boris.unibe.ch/id/eprint/118239

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