Murine Glia Isolation
Murine Glia Isolation - Related Videos
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Dissection and Isolation of Murine Glia from Multiple Central Nervous System Regions
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Cited by 4 •
2020
Here we present a protocol for in vitro isolation of multiple glial cell populations from a mouse CNS. This method allows for the segregation of regional microglia, oligodendrocyte precursor cells, and astrocytes to study the phenotypes of each in a variety of culture systems.
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An In-vitro Preparation of Isolated Enteric Neurons and Glia from the Myenteric Plexus of the Adult Mouse
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Cited by 68 •
2013
We demonstrate a cell culture protocol for the direct study of neuronal and glial components of the enteric nervous system. A neuron/glia mixed culture on coverslips is prepared from the myenteric plexus of adult mouse providing the ability to examine individual neuron and glia function by electrophysiology, immunohistochemical, etc.
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Bilaminar Co-culture of Primary Rat Cortical Neurons and Glia
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Cited by 26 •
2011
Here we provide a protocol for culturing rat cortical neurons in the presence of a glial feeder layer. The cultured neurons establish polarity and create synapses, and can be separated from the glia for use in various applications, such as electrophysiology, calcium imaging, cell survival assays, immunocytochemistry, and RNA/DNA/protein isolation.
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Isolation of Murine Embryonic Hemogenic Endothelial Cells
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Cited by 11 •
2016
Hematopoietic stem and progenitor cells (HSPC) derive from specialized (hemogenic) endothelial cells during development, yet little is known about the process by which some endothelial cells specify to become blood forming. We demonstrate a flow-cytometry based method allowing simultaneous isolation of hemogenic endothelial cells and HSPC from murine embryonic tissues.
08:08
Isolation and Activation of Murine Lymphocytes
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Cited by 56 •
2016
Lymphocytes are the major players in adaptive immune responses. Here, we present a lymphocyte purification protocol to determine the physiological functions of the desired molecules in lymphocyte activation in vitro and in vivo. The described experimental procedures are suitable for comparing functional capacities between control and genetically modified lymphocytes.
FAQs
Enrichment depends on how the dissociated cells are handled after tissue disruption. Selective culture conditions, differences in adhesion, or cell-sorting strategies can favor recovery of particular glial populations from the mixed suspension. The chosen route therefore shapes which cells become available for analysis, making enrichment central to connecting an isolated population with a specific biological question.
Different isolated populations support different neuroscience questions. Astrocytes, microglia, oligodendrocyte-lineage cells, and other glia can be examined in relation to neuroinflammation, myelination, neuron–glia interactions, injury responses, or drug responses. Selecting the relevant population helps researchers create a cellular model aligned with the mechanism or disease process under investigation.
Mechanical and enzymatic dissociation provide alternative ways to convert mouse nervous tissue into a cell suspension before enrichment. They represent the tissue-disruption stage rather than the population-selection stage. After dissociation, researchers can apply selective culture conditions, adhesion-based enrichment, or cell sorting to obtain glial preparations suited to downstream neuroscience experiments.
A typical workflow begins with mouse nervous tissue, proceeds through mechanical or enzymatic dissociation, and produces a suspension containing neural cells. Researchers then enrich desired glial populations using selective culture conditions, adhesion properties, or cell-sorting strategies. The resulting preparation can be used as a controlled cellular model for molecular or functional studies.
Researchers use these preparations when they need to examine glial behavior under controlled conditions rather than study the entire nervous tissue at once. Applications supported by the method include investigating neuroinflammation, myelination, neuron–glia interactions, injury responses, and drug effects. The approach can also help analyze cellular mechanisms relevant to nervous-system disorders.
Isolated glial populations provide tractable systems for studying cellular and molecular mechanisms of brain function and disease. They allow experiments to focus on responses from selected astrocyte, microglial, oligodendrocyte-lineage, or other glial populations. In neuroscience, this can clarify how glial cells participate in inflammation, myelination, injury responses, and interactions with neurons.