title: [Extraction and identification of semen-derived exosomes using PEG6000].
abstract: OBJECTIVE
  To extract and identify semen-derived exosome using PEG6000.
  
  
  METHODS
  Exosomes were extracted from semen specimens from 6 healthy volunteers with step-by-step centrifugations and ultracentrifugation prior to 8% PEG6000 enrichment. The extracted exosomes were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and Western blotting.
  
  
  RESULTS
  The pellets obtained were round or elliptic membrane vesicles 30 to 150 nm in diameter with intact double membranes and contained low electron density material. The pellets expressed CD63, ALIX and TSG101 molecules but not calnexin that was expressed in sperm cells.
  
  
  CONCLUSIONS
  The PEG6000-based method for extraction of exosomes from semen samples facilitates future studies of seminal exosomes.
authors.affiliation: Department of Urology, Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, China.E-mail: yaking91@163.com.
authors.forename: Cheng
authors.initials: C
authors.lastname: Yang
authors.forename: Wen-Bin
authors.initials: WB
authors.lastname: Guo
authors.forename: Wan-Song
authors.initials: WS
authors.lastname: Zhang
authors.forename: Jun
authors.initials: J
authors.lastname: Bian
authors.forename: Jian-Kun
authors.initials: JK
authors.lastname: Yang
authors.forename: Tao
authors.initials: T
authors.lastname: Qi
authors.forename: Chun-Yan
authors.initials: CY
authors.lastname: Wang
authors.forename: Cun-Dong
authors.initials: CD
authors.lastname: Liu
mesh_terms: D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D002498:Centrifugation; D004268:DNA-Binding Proteins / Q000378:metabolism; D056827:Endosomal Sorting Complexes Required for Transport / Q000378:metabolism; D055354:Exosomes / Q000737:chemistry*; D006801:Humans; D008297:Male; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles; D011092:Polyethylene Glycols; D012661:Semen / Q000737:chemistry*; D013094:Spermatozoa; D060149:Tetraspanin 30 / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014461:Ultracentrifugation; D000099177:Tsg101 Protein
chemical_list: D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D011092:Polyethylene Glycols; D060149:Tetraspanin 30; D014157:Transcription Factors; D000099177:Tsg101 Protein; C554586:CD63 protein, human; C117020:PDCD6IP protein, human; C000595215:Polyethylene Glycol 6000