PMID- 35864901
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20230919
IS  - 2331-8325 (Electronic)
IS  - 2331-8325 (Linking)
VI  - 12
IP  - 12
DP  - 2022 Jun 20
TI  - Fluorescent Labeling of Small Extracellular Vesicles (EVs) Isolated from 
      Conditioned Media.
LID - e4447 [pii]
LID - 10.21769/BioProtoc.4447 [doi]
AB  - Extracellular vesicles (EVs), such as exosomes, are produced by all known 
      eukaryotic cells, and constitute essential means of intercellular communication. 
      Recent studies have unraveled the important roles of EVs in migrating to specific 
      sites and cells. Functional studies of EVs using in vivo and in vitro systems 
      require tracking these organelles using fluorescent dyes or, alternatively, 
      transfected and fluorescent-tagged proteins, located either intravesicularly or 
      anchored to the EV bilayer membrane. Due to design simplicity, the fluorescent 
      dye might be a preferred method if the cells are difficult to modify by 
      transfection or when the genetic alteration of the mother cells is not desired. 
      This protocol describes techniques to label cultured cell-derived EVs, using 
      lipophilic DiR [DiIC18(7) 
      (1,1'-Dioctadecyl-3,3,3',3'-Tetramethylindotricarbocyanine Iodide)] fluorophore. 
      This technique can be used to study the cellular uptake and intracellular 
      localization of EVs, and their biodistribution in vivo , which are crucial 
      evaluations of any isolated EVs.
CI  - Copyright (c) 2022 The Authors; exclusive licensee Bio-protocol LLC.
FAU - Santelices, John
AU  - Santelices J
AD  - Department of Microbiology and Cell Science, Institute of Food and Agricultural 
      Sciences, University of Florida, Gainesville, Florida, United States of America.
FAU - Ou, Mark
AU  - Ou M
AD  - Department of Microbiology and Cell Science, Institute of Food and Agricultural 
      Sciences, University of Florida, Gainesville, Florida, United States of America.
FAU - Hui, Winnie W
AU  - Hui WW
AD  - Department of Microbiology and Cell Science, Institute of Food and Agricultural 
      Sciences, University of Florida, Gainesville, Florida, United States of America.
FAU - Maegawa, Gustavo H B
AU  - Maegawa GHB
AD  - Department of Pediatrics, Columbia University Irving Medical Center, New 
      York-Presbyterian Morgan Stanley Children's Hospital, New York, United States of 
      America.
FAU - Edelmann, Mariola J
AU  - Edelmann MJ
AD  - Department of Microbiology and Cell Science, Institute of Food and Agricultural 
      Sciences, University of Florida, Gainesville, Florida, United States of America.
LA  - eng
GR  - R01 AI158749/AI/NIAID NIH HHS/United States
GR  - R03 AI135610/AI/NIAID NIH HHS/United States
GR  - R21 NS113649/NS/NINDS NIH HHS/United States
PT  - Journal Article
DEP - 20220620
PL  - United States
TA  - Bio Protoc
JT  - Bio-protocol
JID - 101635102
PMC - PMC9257841
OTO - NOTNLM
OT  - DiR
OT  - Exosomes
OT  - Extracellular vesicles
OT  - Labeling
OT  - Lipophilic dye
COIS- Competing interests The authors have declared that no competing interests exist.
EDAT- 2022/07/23 06:00
MHDA- 2022/07/23 06:01
PMCR- 2022/06/20
CRDT- 2022/07/22 02:16
PHST- 2022/02/01 00:00 [received]
PHST- 2022/04/24 00:00 [revised]
PHST- 2022/04/26 00:00 [accepted]
PHST- 2022/07/22 02:16 [entrez]
PHST- 2022/07/23 06:00 [pubmed]
PHST- 2022/07/23 06:01 [medline]
PHST- 2022/06/20 00:00 [pmc-release]
AID - e4447 [pii]
AID - 4447 [pii]
AID - 10.21769/BioProtoc.4447 [doi]
PST - epublish
SO  - Bio Protoc. 2022 Jun 20;12(12):e4447. doi: 10.21769/BioProtoc.4447. eCollection 
      2022 Jun 20.