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Article . 2016
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Isolation and Flow Cytometric Analysis of Immune Cells from the Ischemic Mouse Brain

Authors: Gesa Weise; Daniel-Christoph Wagner; Johannes Boltze; Karoline Möller; Claudia Pösel;

Isolation and Flow Cytometric Analysis of Immune Cells from the Ischemic Mouse Brain

Abstract

Ischemic stroke initiates a robust inflammatory response that starts in the intravascular compartment and involves rapid activation of brain resident cells. A key mechanism of this inflammatory response is the migration of circulating immune cells to the ischemic brain facilitated by chemokine release and increased endothelial adhesion molecule expression. Brain-invading leukocytes are well-known contributing to early-stage secondary ischemic injury, but their significance for the termination of inflammation and later brain repair has only recently been noticed. Here, a simple protocol for the efficient isolation of immune cells from the ischemic mouse brain is provided. After transcardial perfusion, brain hemispheres are dissected and mechanically dissociated. Enzymatic digestion with Liberase is followed by density gradient (such as Percoll) centrifugation to remove myelin and cell debris. One major advantage of this protocol is the single-layer density gradient procedure which does not require time-consuming preparation of gradients and can be reliably performed. The approach yields highly reproducible cell counts per brain hemisphere and allows for measuring several flow cytometry panels in one biological replicate. Phenotypic characterization and quantification of brain-invading leukocytes after experimental stroke may contribute to a better understanding of their multifaceted roles in ischemic injury and repair.

Country
Germany
Subjects by Vocabulary

Microsoft Academic Graph classification: Pathology Chemokine Cell Myelin Stroke medicine.diagnostic_test biology medicine.anatomical_structure medicine.symptom medicine.medical_specialty Inflammation Flow cytometry Immune system medicine business.industry medicine.disease biology.protein business Percoll

Keywords

Male, General Chemical Engineering, Cell Count, Cell Separation, cell phenotyping, characterization, Myelin Sheath, Ischemic stroke, Dissection, General Neuroscience, Brain, Infarction, Middle Cerebral Artery, Stroke, Encephalitis, Microglia, leukocytes, Immunology, General Biochemistry, Genetics and Molecular Biology, cell isolation, immune cells, Centrifugation, Density Gradient, Animals, brain inflammation, General Immunology and Microbiology, flow cytometry, Suture Techniques, quantification, Mice, Inbred C57BL, Disease Models, Animal, repair, Issue 108

30 references, page 1 of 3

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    Top 10%
    influence
    This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    Top 10%
    impulse
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    Top 10%
  • citations
    This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    61
    popularity
    This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
    Top 10%
    influence
    This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    Top 10%
    impulse
    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
    Top 10%
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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
61
Top 10%
Top 10%
Top 10%
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bronze
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Neuroinformatics
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