title: [Quantification and size distribution of 24-hour urinary extracellular vesicles from healthy adults].
abstract: OBJECTIVE
  To analyze the quantity and size distribution of 24-hour urinary extracellular vesicles (uEVs) from healthy adults.
  
  
  METHODS
  The 24-hour uEVs from 9 healthy adults were isolated by hydrostatic filtration dialysis (HFD). The effectiveness of uEVs enrichment was evaluated using Western blotting and transmission electron microscopy (TEM). The quantity and size distribution of the uEVs was analyzed with BCA protein quantification, TEM, and nanoparticle tracking analysis (NTA).
  
  
  RESULTS
  uEVs with different sizes and morphologies were observed under TEM. Western blotting confirmed the expression of TSG101 in all the uEV fractions from the 9 donors, ranging from 132.50 to 760.70 ng/mL. NTA results showed that the number of 24-hour uEVs amount ranged from 3.56 × 10¹² particles to 5.12 × 10¹² particles, with a CV of 14.23%. The proportion of the vesicles with a diameter <40 nm was 0.04%-0.69% with a number range of (1.80-26.49)× 10⁹ particles; the proportion of vesicles with a diameter of 40-100 nm (which is consistent with the size of exosomes)was 22.07%-42.08% with a number range of (1.00-1.77)× 10¹² particles. The proportion of vesicles with a diameter of 100-1000 nm (consistent with the size of microvesicles) was 57.88%-77.85% with a number range of (2.09-3.86)× 10¹² particles.
  
  
  CONCLUSIONS
  The established HFD method allows efficient and convenient isolation of uEVs from a large amount of urine samples. The 24-hour uEVs from healthy adults show narrow differences between individuals and thus can be an ideal source of samples for relevant studies.
authors.affiliation: Institute of Nephrology and Urology, Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, China.E-mail: hanfeilin@hotmail.com.
authors.forename: Hanfei
authors.initials: H
authors.lastname: Lin
authors.forename: Xinyu
authors.initials: X
authors.lastname: Liu
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Xu
authors.forename: Luca Musante
authors.initials: LM
authors.lastname: Luca Musante
authors.forename: Harry Holthofer
authors.initials: HH
authors.lastname: Harry Holthofer
authors.forename: Hequn
authors.initials: H
authors.lastname: Zou
mesh_terms: D000328:Adult; D015153:Blotting, Western; D055252:Cell-Derived Microparticles; D055354:Exosomes; D000067128:Extracellular Vesicles; D006801:Humans; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles; D014556:Urine