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Different EV enrichment methods suitable for clinical settings yield different
subpopulations of urinary extracellular vesicles from human samples
#MMPMID26895490
Royo F
; Zuņiga-Garcia P
; Sanchez-Mosquera P
; Egia A
; Perez A
; Loizaga A
; Arceo R
; Lacasa I
; Rabade A
; Arrieta E
; Bilbao R
; Unda M
; Carracedo A
; Falcon-Perez JM
J Extracell Vesicles
2016[]; 5
(?): 29497
PMID26895490
show ga
Urine sample analysis is irreplaceable as a non-invasive method for disease
diagnosis and follow-up. However, in urine samples, non-degraded protein and RNA
may be only found in urinary extracellular vesicles (uEVs). In recent years,
various methods of uEV enrichment using low volumes of urine and unsophisticated
equipment have been developed, with variable success. We compared the results of
the differential ultracentrifugation procedure with 4 of these methods. The
methods tested were a lectin-based purification, Exoquick (System Biosciences),
Total Exosome Isolation from Invitrogen and an in-house modified procedure
employing the Exosomal RNA Kit from Norgen Biotek Corp. The analysis of selected
gene transcripts and protein markers of extracellular vesicles (EVs) revealed
that each method isolates a different mixture of uEV protein markers. In our
conditions, the extraction with Norgen's reagent achieved the best performance in
terms of gene transcript and protein detection and reproducibility. By using this
method, we were able to detect alterations of EVs protein markers in urine
samples from prostate cancer adenoma patients. Taken together, our results show
that the isolation of uEVs is feasible from small volumes of urine and avoiding
ultracentrifugation, making easier the analysis in a clinical facility. However,
caution should be taken in the selection of the enrichment method since they have
a differential affinity for protein uEVs markers and by extension for different
subpopulation of EVs.