Characterization and isolation of mouse primary microglia by density gradient centrifugation

Jessica C. Stark, Euan Wallace, Rebecca Lim, Bryan Leaw

Research output: Contribution to journalArticleOtherpeer-review

Abstract

Microglia, the resident immune cells in the brain, are the first responders to inflammation or injury in the central nervous system. Recent research has revealed microglia to be dynamic, capable of assuming both pro-inflammatory and anti-inflammatory phenotypes. Both M1 (pro-inflammatory) and M2 (pro-reparative) phenotypes play an important role in neuroinflammatory conditions such as perinatal brain injury, and exhibit differing functions in response to certain environmental stimuli. The modulation of microglial activation has been noted to confer neuroprotection thus suggesting microglia may have therapeutic potential in brain injury. However, more research is required to better understand the role of microglia in disease, and this protocol facilitates that. The protocol described below combines a density gradient centrifugation process to reduce cellular debris, with magnetic separation, producing a highly pure sample of primary microglial cells that can be used for in vitro experimentation, without the need for 2-3 weeks culturing. Additionally, the characterization steps yield robust functional data about microglia, aiding studies to better our understanding of the polarization and priming of these cells, which has strong implications in the field of regenerative medicine.

Original languageEnglish
Article numbere57065
Number of pages9
JournalJournal of Visualized Experiments
Volume2018
Issue number132
DOIs
Publication statusPublished - 16 Feb 2018

Keywords

  • Animal model
  • Cell culture
  • Inflammation
  • Isolation
  • Issue 132
  • Microglia
  • Mouse
  • Neonatal
  • Neuroscience
  • Perinatal brain injury
  • Primary cell

Cite this

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title = "Characterization and isolation of mouse primary microglia by density gradient centrifugation",
abstract = "Microglia, the resident immune cells in the brain, are the first responders to inflammation or injury in the central nervous system. Recent research has revealed microglia to be dynamic, capable of assuming both pro-inflammatory and anti-inflammatory phenotypes. Both M1 (pro-inflammatory) and M2 (pro-reparative) phenotypes play an important role in neuroinflammatory conditions such as perinatal brain injury, and exhibit differing functions in response to certain environmental stimuli. The modulation of microglial activation has been noted to confer neuroprotection thus suggesting microglia may have therapeutic potential in brain injury. However, more research is required to better understand the role of microglia in disease, and this protocol facilitates that. The protocol described below combines a density gradient centrifugation process to reduce cellular debris, with magnetic separation, producing a highly pure sample of primary microglial cells that can be used for in vitro experimentation, without the need for 2-3 weeks culturing. Additionally, the characterization steps yield robust functional data about microglia, aiding studies to better our understanding of the polarization and priming of these cells, which has strong implications in the field of regenerative medicine.",
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Characterization and isolation of mouse primary microglia by density gradient centrifugation. / Stark, Jessica C.; Wallace, Euan; Lim, Rebecca; Leaw, Bryan.

In: Journal of Visualized Experiments, Vol. 2018, No. 132, e57065, 16.02.2018.

Research output: Contribution to journalArticleOtherpeer-review

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