PMID- 32568235
OWN - NLM
STAT- MEDLINE
DCOM- 20201014
LR  - 20201014
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 160
DP  - 2020 Jun 2
TI  - Characterization of Immune Cell-derived Extracellular Vesicles and Studying 
      Functional Impact on Cell Environment.
LID - 10.3791/60118 [doi]
AB  - The neuroinflammatory state of the central nervous system (CNS) plays a key role 
      in physiological and pathological conditions. Microglia, the resident immune 
      cells in the brain, and sometimes the infiltrating bone marrow-derived 
      macrophages (BMDMs), regulate the inflammatory profile of their microenvironment 
      in the CNS. It is now accepted that the extracellular vesicle (EV) populations 
      from immune cells act as immune mediators. Thus, their collection and isolation 
      are important to identify their contents but also evaluate their biological 
      effects on recipient cells. The present data highlight chronological requirements 
      for EV isolation from microglia cells or blood macrophages including the 
      ultracentrifugation and size-exclusion chromatography (SEC) steps. A non-targeted 
      proteomic analysis permitted the validation of protein signatures as EV markers 
      and characterized the biologically active EV contents. Microglia-derived EVs were 
      also functionally used on primary culture of neurons to assess their importance 
      as immune mediators in the neurite outgrowth. The results showed that 
      microglia-derived EVs contribute to facilitate the neurite outgrowth in vitro. In 
      parallel, blood macrophage-derived EVs were functionally used as immune mediators 
      in spheroid cultures of C6 glioma cells, the results showing that these EVs 
      control the glioma cell invasion in vitro. This report highlights the possibility 
      to evaluate the EV-mediated immune cell functions but also understand the 
      molecular bases of such a communication. This deciphering could promote the use 
      of natural vesicles and/or the in vitro preparation of therapeutic vesicles in 
      order to mimic immune properties in the microenvironment of CNS pathologies.
FAU - Lemaire, Quentin
AU  - Lemaire Q
AD  - U1192-Laboratoire Proteomique, Reponse Inflammatoire et Spectrometrie de Masse 
      (PRISM), Univ. Lille, INSERM.
FAU - Duhamel, Marie
AU  - Duhamel M
AD  - U1192-Laboratoire Proteomique, Reponse Inflammatoire et Spectrometrie de Masse 
      (PRISM), Univ. Lille, INSERM.
FAU - Raffo-Romero, Antonella
AU  - Raffo-Romero A
AD  - U1192-Laboratoire Proteomique, Reponse Inflammatoire et Spectrometrie de Masse 
      (PRISM), Univ. Lille, INSERM.
FAU - Salzet, Michel
AU  - Salzet M
AD  - U1192-Laboratoire Proteomique, Reponse Inflammatoire et Spectrometrie de Masse 
      (PRISM), Univ. Lille, INSERM.
FAU - Lefebvre, Christophe
AU  - Lefebvre C
AD  - U1192-Laboratoire Proteomique, Reponse Inflammatoire et Spectrometrie de Masse 
      (PRISM), Univ. Lille, INSERM; christophe.lefebvre@univ-lille.fr.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20200602
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
SB  - IM
MH  - Animals
MH  - Brain/immunology/pathology
MH  - Cell Line, Tumor
MH  - Macrophages/*cytology
MH  - Microglia/cytology
MH  - Proteomics
MH  - Rats
MH  - Tumor Microenvironment
EDAT- 2020/06/23 06:00
MHDA- 2020/10/21 06:00
CRDT- 2020/06/23 06:00
PHST- 2020/06/23 06:00 [entrez]
PHST- 2020/06/23 06:00 [pubmed]
PHST- 2020/10/21 06:00 [medline]
AID - 10.3791/60118 [doi]
PST - epublish
SO  - J Vis Exp. 2020 Jun 2;(160). doi: 10.3791/60118.