ArticleJournal of extracellular vesicles2021
Urinary extracellular vesicles: Assessment of pre-analytical variables and development of a quality control with focus on transcriptomic biomarker research.
Article in Journal of extracellular vesicles, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 41 papers, 1 of them a synthesis that pooled it.
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Who cites it
41 citing papers in PubMed, 1 synthesis or guideline pooled it, 56 citations in OpenAlex.
- Different storage and freezing protocols for extracellular vesicles: a systematic review.Stem cell research & therapy · 2024Pooled it
- Sex differences in urinary extracellular vesicles originating from the genitourinary system in health and disease.American journal of physiology. Renal physiology · 2026Review
- Review
- A high-throughput, end-to-end pipeline for extracellular miRNA biomarker discovery from human biofluids.Cell reports methods · 2026Article
- Urinary Extracellular Vesicle-Derived miRNAs as Regulators and Biomarkers in Diabetic Kidney Disease.International journal of molecular sciences · 2026Review
- Urinary Extracellular Vesicles Biomarkers in CKD: Clinical Laboratory Translation.Diagnostics (Basel, Switzerland) · 2026Review
- Development and validation of a noninvasive machine learning model using urinary extracellular vesicle physical parameters for prostate cancer diagnosis.Scientific reports · 2026Article
- Exosomes as Disease-Informed Nanoplatforms for Pulmonary Fibrosis: From Pathogenic Signaling to Precision Diagnosis and Therapy.Pharmaceutics · 2026Review
- Oral natural extracellular vesicles for biomedical applications: Advances and clinical perspectives.Journal of advanced research · 2026Review
- From feasibility to translational pathways: a bibliometric and knowledge-mapping analysis of urine-based liquid biopsy in urologic cancers (2015-2025).Journal of translational medicine · 2026Review
- Article
- Urine-based PSMA detection for noninvasive prostate cancer diagnosis: recent advances, translational challenges, and future perspectives.Frontiers in oncology · 2026Review
- Quantification of EV-associated miRNA in liquid biopsies for biomarker signature development.Extracellular vesicles and circulating nucleic acids · 2026Review
- Unveiling Heterogeneity: Innovations and Challenges in Single-Vesicle Analysis for Clinical Translation.Journal of extracellular vesicles · 2025Article
- Selected miRNAs in Urinary Extracellular Vesicles Show Promise for Early and Specific Diagnostics of Diabetic Kidney Disease.Journal of extracellular biology · 2025Article
- Exo-hydrogel therapy: a revolutionary approach to managing diabetic complications.Journal of nanobiotechnology · 2025Review
- Single urinary extracellular vesicle proteomics identifies complement receptor CD35 as a biomarker for sepsis-associated acute kidney injury.Nature communications · 2025Article
- Roadblocks of Urinary EV Biomarkers: Moving Toward the Clinic.Journal of extracellular vesicles · 2025Review
- Differentiation of high risk prostate cancer with a facile urinary exosome detection workflow.iScience · 2025Article
- Extracellular vesicles in female reproduction: from basic research to application.Animal reproduction · 2025Review
Corrections and comments
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Authors and funding
15 authors at 4 institutions in 5 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Urinary extracellular vesicles (uEV) are a topical source of non-invasive biomarkers for health and diseases of the urogenital system. However, several challenges have become evident in the standardization of uEV pipelines from collection of urine to biomarker analysis. Here, we studied the effect of pre-analytical variables and developed means of quality control for uEV isolates to be used in transcriptomic biomarker research. We included urine samples from healthy controls and individuals with type 1 or type 2 diabetes and normo-, micro- or macroalbuminuria and isolated uEV by ultracentrifugation. We studied the effect of storage temperature (-20°C vs. -80°C), time (up to 4 years) and storage format (urine or isolated uEV) on quality of uEV by nanoparticle tracking analysis, electron microscopy, Western blotting and qPCR. Urinary EV RNA was compared in terms of quantity, quality, and by mRNA or miRNA sequencing. To study the stability of miRNA levels in samples isolated by different methods, we created and tested a list of miRNAs commonly enriched in uEV isolates. uEV and their transcriptome were preserved in urine or as isolated uEV even after long-term storage at -80°C. However, storage at -20°C degraded particularly the GC-rich part of the transcriptome and EV protein markers. Transcriptome was preserved in RNA samples extracted with and without DNAse, but read distributions still showed some differences in e.g. intergenic and intronic reads. MiRNAs commonly enriched in uEV isolates were stable and concordant between different EV isolation methods. Analysis of never frozen uEV helped to identify surface characteristics of particles by EM. In addition to uEV, qPCR assays demonstrated that uEV isolates commonly contained polyoma viruses. Based on our results, we present recommendations how to store and handle uEV isolates for transcriptomics studies that may help to expedite standardization of the EV biomarker field.
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