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JoVE Journal
Immunology and Infection
Isolating Central Nervous System Tissues and Associated Meninges for the Downstream Analysis of I...
Isolating Central Nervous System Tissues and Associated Meninges for the Downstream Analysis of I...
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Isolating Central Nervous System Tissues and Associated Meninges for the Downstream Analysis of Immune cells

Isolating Central Nervous System Tissues and Associated Meninges for the Downstream Analysis of Immune cells

Full Text
10,939 Views
09:35 min
May 19, 2020

DOI: 10.3791/61166-v

Krista D. DiSano1, Michael R. Linzey1,2, Nora C. Welsh1,2, Joshua S. Meier1, Andrew R. Pachner1, Francesca Gilli1

1Department of Neurology, Geisel School of Medicine,Dartmouth-Hitchcock Medical Center, 2Program in Experimental and Molecular Medicine,Dartmouth College

This paper presents two optimized protocols for examining resident and peripherally derived immune cells within the central nervous system, including the brain, spinal cord, and meninges. Each of these protocols helps to ascertain the function and composition of the cells occupying these compartments under steady state and inflammatory conditions.

Rapid isolation of CNS tissues including the meninges allows for a comprehensive analysis of immune cell phenotype, function, and localization in homeostatic and pathogenic conditions. The two demonstrated methods allow for the quick extraction of CNS tissues with the meninges, ideal for downstream analysis using single cell techniques and histological methods. Although the representative results focus on the analysis of immune cells, these two methods can be used to analyze CNS resident cells, including microglia, astrocytes, pericytes, endothelial cells, stromal cells, and neurons.

Begin by isolating brain and spinal cord samples from euthanized mice. After removing the head, make a midline incision in the skin and flip it over the eyes to free the skull. Cut at the nasal bone to release the mandible from the skull.

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