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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

Article DOI: 10.3791/51005-v • 09:12 min • January 30th, 2014
January 30th, 2014 •
1, 1, 1
1Department of Neuroscience, Georgetown University Medical Center

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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).

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Isolation Cortical Microglia Immunophenotype Functionality Murine Neonates CNS Tissue Investigative Tool Microglial Biology Ex Vivo Procedure Mechanical Agitation Rotary Shaker Highly Pure Morphological Characteristics Functional Characteristics Quiescent Microglia Nonpathological Conditions In Vivo Preservation Biochemical Functionality Morphological Changes Nuclear Translocation P65 Subunit Of NF-κB (p65) Secretion Proinflammatory Cytokine Tumor Necrosis Factor-α (TNF-α) Lipopolysaccharide (LPS) Pam3CSK4 (Pam) Challenges
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