» Articles » PMID: 23498743

Purification of Exosome-like Vesicles from Urine

Overview
Journal Methods Enzymol
Specialty Biochemistry
Date 2013 Mar 19
PMID 23498743
Citations 23
Authors
Affiliations
Soon will be listed here.
Abstract

Urinary exosome-like vesicles (ELVs), 20-200nm membrane-bound particles shed by renal epithelium, have been shown to interact with the primary cilia of distant epithelial cells of the nephron. These ELVs are emerging as an important source of protein, mRNA, and miRNA biomarkers to monitor renal disease. However, purification of ELVs is compromised by the presence of large amounts of the urinary protein Tamm-Horsfall Protein (THP). THP molecules oligomerize into long, double-helical strands several microns long. These linear assemblies form a three-dimensional gel which traps and sequesters ELVs in any centrifugation-based protocol. Here, we present a purification protocol that separates ELVs from THP and divides urinary ELVs into three distinct populations.

Citing Articles

High-throughput surface epitope immunoaffinity isolation of extracellular vesicles and downstream analysis.

Khanabdali R, Mandrekar M, Grygiel R, Vo P, Palma C, Nikseresht S Biol Methods Protoc. 2024; 9(1):bpae032.

PMID: 39070184 PMC: 11272960. DOI: 10.1093/biomethods/bpae032.


Exosomes promote pre-metastatic niche formation in colorectal cancer.

Si G, Chen X, Li Y, Yuan X Heliyon. 2024; 10(6):e27572.

PMID: 38509970 PMC: 10950591. DOI: 10.1016/j.heliyon.2024.e27572.


Urinary Exosomal MicroRNAs as Biomarkers for Obesity-Associated Chronic Kidney Disease.

Earle A, Bessonny M, Benito J, Huang K, Parker H, Tyler E J Clin Med. 2022; 11(18).

PMID: 36142918 PMC: 9502686. DOI: 10.3390/jcm11185271.


Using human urinary extracellular vesicles to study physiological and pathophysiological states and regulation of the sodium chloride cotransporter.

Wu A, Wolley M, Fenton R, Stowasser M Front Endocrinol (Lausanne). 2022; 13:981317.

PMID: 36105401 PMC: 9465297. DOI: 10.3389/fendo.2022.981317.


Isolation of extra-cellular vesicles in the context of pancreatic adenocarcinomas: Addition of one stringent filtration step improves recovery of specific microRNAs.

Xu Y, Xu X, Bhandari K, Gin A, Rao C, Morris K PLoS One. 2021; 16(11):e0259563.

PMID: 34784377 PMC: 8594802. DOI: 10.1371/journal.pone.0259563.


References
1.
Miranda K, Bond D, McKee M, Skog J, Paunescu T, da Silva N . Nucleic acids within urinary exosomes/microvesicles are potential biomarkers for renal disease. Kidney Int. 2010; 78(2):191-9. PMC: 4451567. DOI: 10.1038/ki.2010.106. View

2.
Hogan M, Manganelli L, Woollard J, Masyuk A, Masyuk T, Tammachote R . Characterization of PKD protein-positive exosome-like vesicles. J Am Soc Nephrol. 2009; 20(2):278-88. PMC: 2637052. DOI: 10.1681/ASN.2008060564. View

3.
Pieper R, Gatlin C, McGrath A, Makusky A, Mondal M, Seonarain M . Characterization of the human urinary proteome: a method for high-resolution display of urinary proteins on two-dimensional electrophoresis gels with a yield of nearly 1400 distinct protein spots. Proteomics. 2004; 4(4):1159-74. DOI: 10.1002/pmic.200300661. View

4.
Zhou H, Yuen P, Pisitkun T, Gonzales P, Yasuda H, Dear J . Collection, storage, preservation, and normalization of human urinary exosomes for biomarker discovery. Kidney Int. 2006; 69(8):1471-6. PMC: 2276656. DOI: 10.1038/sj.ki.5000273. View

5.
Parolini I, Federici C, Raggi C, Lugini L, Palleschi S, De Milito A . Microenvironmental pH is a key factor for exosome traffic in tumor cells. J Biol Chem. 2009; 284(49):34211-22. PMC: 2797191. DOI: 10.1074/jbc.M109.041152. View