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Magnetic Isolation of Microglial Cells from Neonate Mouse for Primary Cell Cultures
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Isolation and Phenotyping of Adult Mouse Microglial Cells

Kathleen Grabert1, Barry W McColl2,3

  • 1The Roslin Institute and R(D)SVS, University of Edinburgh, Edinburgh, UK. Kathleen.Grabert@roslin.ed.ac.uk.

Methods in Molecular Biology (Clifton, N.J.)
|May 16, 2018
PubMed

Insights

This study presents a refined protocol for isolating pure microglia from adult mouse brains. This method yields viable cells for various functional and molecular analyses, advancing central nervous system research.

Area of Science:

  • Neuroscience
  • Immunology
  • Cell Biology

Background:

  • Microglia are key immune cells in the central nervous system (CNS) involved in development, homeostasis, and inflammation.
  • Existing protocols for microglia isolation are often limited, particularly for adult mouse brains.
  • Pure microglia populations are essential for studying their specific functions and phenotypes.

Purpose of the Study:

  • To describe a refined protocol for the efficient isolation of pure, viable microglia from adult mouse brains.
  • To enable downstream functional and molecular analyses of microglia without contamination from other cell types.

Main Methods:

  • Enzymatic and mechanical dissociation of adult mouse brain tissue.
  • Myelin removal using Percoll density gradient centrifugation.
  • Immunomagnetic selection for microglia isolation.

Main Results:

  • The protocol yields a pure population of microglia from adult mouse brains.
  • Isolated microglia are functionally viable for diverse applications.
  • The method is suitable for various downstream analyses including cell culture, flow cytometry, phagocytosis assays, and transcriptome profiling.

Conclusions:

  • This refined protocol offers a robust method for obtaining pure, viable adult mouse microglia.
  • The isolated cells support a wide range of experimental applications, facilitating CNS research.
  • This technique advances the study of microglial roles in brain health and disease.
Keywords:
CD11b immunomagnetic beadsEnzyme digestionMicrogliaPercoll gradient

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