PMID- 32111925
OWN - NLM
STAT- MEDLINE
DCOM- 20201109
LR  - 20240423
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 10
IP  - 1
DP  - 2020 Feb 28
TI  - Rigorous characterization of urinary extracellular vesicles (uEVs) in the low 
      centrifugation pellet - a neglected source for uEVs.
PG  - 3701
LID - 10.1038/s41598-020-60619-w [doi]
LID - 3701
AB  - Urinary extracellular vesicles (uEVs) provide bio-markers for kidney and 
      urogenital diseases. Centrifugation is the most common method used to enrich 
      uEVs. However, a majority of studies to date have focused on the 
      ultracentrifugation pellet, potentially losing a novel source of important 
      biomarkers that could be obtained at lower centrifugation. Thus, the aim of this 
      study is to rigorously characterize for the first time uEVs in the low speed 
      pellet and determine the minimal volume of urine required for proteomic analysis 
      (>/=9.0 mL urine) and gene ontology classification identified 75% of the protein as 
      extracellular exosomes. Cryo-Transmission Electron Microscopy (>/=3.0 mL urine) 
      provided evidence of a heterogeneous population of EVs for size and morphology 
      independent of uromodulin filaments. Western blot detected several specific uEV 
      kidney and EV markers (>/=4.5 mL urine per lane). microRNAs quantification by qPCR 
      was possible with urine volume as low as 0.5 mL. Particle enumeration with 
      tunable resistive pulse sensing, nano particles tracking analysis and single EV 
      high throughput imaging flow cytometry are possible starting from 0.5 and 3.0 mL 
      of urine respectively. This work characterizes a neglected source of uEVs and 
      provides guidance with regard to volume of urine necessary to carry out 
      multi-omic studies and reveals novel aspects of uEV analysis such as 
      autofluorescence of podocyte origin.
FAU - Musante, Luca
AU  - Musante L
AD  - Division of Nephrology and Center for Immunity, Inflammation and Regenerative 
      Medicine, Department of Medicine, University of Virginia, Charlottesville, VA, 
      USA.
FAU - Bontha, Sai Vineela
AU  - Bontha SV
AD  - Transplant Research Institute, James D. Eason Transplant Institute, School of 
      Medicine, University of Tennessee Health Science Center, Memphis, TN, USA.
FAU - La Salvia, Sabrina
AU  - La Salvia S
AD  - Division of Nephrology and Center for Immunity, Inflammation and Regenerative 
      Medicine, Department of Medicine, University of Virginia, Charlottesville, VA, 
      USA.
FAU - Fernandez-Pineros, Angela
AU  - Fernandez-Pineros A
AD  - Transplant Research Institute, James D. Eason Transplant Institute, School of 
      Medicine, University of Tennessee Health Science Center, Memphis, TN, USA.
FAU - Lannigan, Joanne
AU  - Lannigan J
AD  - School of Medicine, Flow Cytometry Core, University of Virginia, Charlottesville, 
      VA, USA.
FAU - Le, Thu H
AU  - Le TH
AD  - Department of Medicine, Division of Nephrology, University of Rochester Medical 
      Center, Rochester, NY, USA.
FAU - Mas, Valeria
AU  - Mas V
AD  - Transplant Research Institute, James D. Eason Transplant Institute, School of 
      Medicine, University of Tennessee Health Science Center, Memphis, TN, USA.
FAU - Erdbrugger, Uta
AU  - Erdbrugger U
AD  - Division of Nephrology and Center for Immunity, Inflammation and Regenerative 
      Medicine, Department of Medicine, University of Virginia, Charlottesville, VA, 
      USA. ue2u@virginia.edu.
LA  - eng
GR  - K23 HL126101/HL/NHLBI NIH HHS/United States
GR  - S10 RR025694/RR/NCRR NIH HHS/United States
GR  - S10 RR031633/RR/NCRR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20200228
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Biomarkers)
SB  - IM
MH  - Adult
MH  - Biomarkers/urine
MH  - Cryoelectron Microscopy
MH  - Extracellular Vesicles/*metabolism/ultrastructure
MH  - Female
MH  - Female Urogenital Diseases/pathology/*urine
MH  - Humans
MH  - Kidney/*metabolism/pathology
MH  - Male
MH  - Male Urogenital Diseases/pathology/*urine
MH  - Middle Aged
MH  - Proteomics
MH  - Ultracentrifugation
PMC - PMC7048852
COIS- The authors declare no competing interests.
EDAT- 2020/03/01 06:00
MHDA- 2020/11/11 06:00
PMCR- 2020/02/28
CRDT- 2020/03/01 06:00
PHST- 2019/11/26 00:00 [received]
PHST- 2020/01/30 00:00 [accepted]
PHST- 2020/03/01 06:00 [entrez]
PHST- 2020/03/01 06:00 [pubmed]
PHST- 2020/11/11 06:00 [medline]
PHST- 2020/02/28 00:00 [pmc-release]
AID - 10.1038/s41598-020-60619-w [pii]
AID - 60619 [pii]
AID - 10.1038/s41598-020-60619-w [doi]
PST - epublish
SO  - Sci Rep. 2020 Feb 28;10(1):3701. doi: 10.1038/s41598-020-60619-w.