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Immunology and Infection
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Isolation of Cortical Microglia with Preserved Immunophenotype and Functionality From Murine Neonates
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Immunology and Infection
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JoVE Journal Immunology and Infection
Isolation of Cortical Microglia with Preserved Immunophenotype and Functionality From Murine Neonates

Isolation of Cortical Microglia with Preserved Immunophenotype and Functionality From Murine Neonates

DOI:
10.3791/51005-v

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09:12 min

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January 30, 2014

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, ,
1Department of Neuroscience,Georgetown University Medical Center

Chapters

  • 00:05Title
  • 01:19Preparation
  • 02:23Sample Collection
  • 04:31Growing Cortical Cells
  • 05:56Isolation of Microglia from Mixed Glial Culture
  • 07:30Results: Preserved Immunophenotype and Functionality in Isolated Microglia
  • 08:46Conclusion

Summary

Automatic Translation

Automatic Translation

One key to successful investigation of microglial biology is the preservation of microglial immunofunction ex vivo during isolation from CNS tissue. Isolating microglia via rotary shaking results in highly pure and immunofunctional cell cultures as assessed by fluorescent imaging, immunocytochemistry, and ELISA following microglia activation with the proinflammatory stimuli lipopolysaccharide (LPS) and Pam3CSK4 (Pam).

Tags

Keywords: Cortical MicrogliaMurine NeonatesMicroglia IsolationImmunophenotypeFunctionalityMechanical AgitationRotary ShakerQuiescent MicrogliaNF-κBTNF-αLipopolysaccharidePam3CSK4

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