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Method Article
Miriam Mecha
Guaza´s Lab, Cajal Institute, Madrid
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
Primary glial cell cultures are the most commonly used in vitro model for neurobiological studies. However, the lack of an easy and fast protocol for the isolation and culture of these cells leads to long incubating times with the use of high number of rat brains. Here, we describe a quick protocol based on mixed glial culture for the highly-enriched isolation of microglia, astrocyte and oligodendrocyte cells. The procedure is suitable for beginners as it makes available the easier way to obtain the nervous tissue, the evolution of the mixed culture throughout the time, and the final results after the isolation of each cell type. More than 20x106 cells per rat brain can be obtained using this protocol, with no more than 11 days of incubation. This facilitates the use of the primary culture as a tool for researchers to study the development, properties and functions of glial cells in vitro.
Keywords
cell culture, microglia, astrocytes, oligodendrocytes
Figure 1
Figure 2
Figure 3
The authors declare no competing financial interests
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.mov
Supplementary video 1 Dissection Method for Primary Cell Culture
- supplement0.mov
Supplementary Video 2 Evolution in time of the Mixed Glial Culture by phase contrast
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CURRENT STATUS: Posted
Version 1
Posted 10 Mar, 2011
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Biological techniques
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Miriam Mecha
Guaza´s Lab, Cajal Institute, Madrid
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 10 Mar, 2011
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Subject Areas
Biological techniques
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
Primary glial cell cultures are the most commonly used in vitro model for neurobiological studies. However, the lack of an easy and fast protocol for the isolation and culture of these cells leads to long incubating times with the use of high number of rat brains. Here, we describe a quick protocol based on mixed glial culture for the highly-enriched isolation of microglia, astrocyte and oligodendrocyte cells. The procedure is suitable for beginners as it makes available the easier way to obtain the nervous tissue, the evolution of the mixed culture throughout the time, and the final results after the isolation of each cell type. More than 20x106 cells per rat brain can be obtained using this protocol, with no more than 11 days of incubation. This facilitates the use of the primary culture as a tool for researchers to study the development, properties and functions of glial cells in vitro.
Figure 1
Figure 2
Figure 3
The authors declare no competing financial interests
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.mov
Supplementary video 1 Dissection Method for Primary Cell Culture
- supplement0.mov
Supplementary Video 2 Evolution in time of the Mixed Glial Culture by phase contrast
Loading...