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Generation of Microglia Cultures and Mixed Glial Culture
Published: Nov 5, 2011 DOI: 10.21769/BioProtoc.149 Views: 19402
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Abstract
Primary rodent microglia-enriched cultures are the most popular model to study microglial biology in vitro and to explore immune signaling pathways. Mixed glial cultures that contain microglia and astroglia are very useful for investigating the precise mechanisms of microglia-astroglia interaction during immune reaction. This protocol has been developed and improved over the years by various researchers in Dr. Hong’s lab, especially Dr. Bin Liu.
Materials and Reagents
- Poly-D-lysine (Sigma-Aldrich, catalog number: P7280 )
- DMEM/F12 (Life Technologies, InvitrogenTM, catalog number: 11330-032 )
- D-Glucose
- Sterile water
- Sterile PBS
- Heat-inactivated fetal bovine serum (FBS) (Life Technologies, Gibco®, catalog number: 16000-044 )
- Non-essential amino acids (100 ml) (Life Technologies, Gibco®, catalog number: 11140-050 )
- Sodium pyruvate (100 ml) (Sigma-Aldrich, catalog number: S8636 )
- 200 mM L-glutamine (100 ml) (Life Technologies, Gibco®, catalog number: 25030-081 )
- Penicillin/streptomycin (100 ml) (Sigma-Aldrich, catalog number: P0781 )
- DMEM/F12-based culture medium (see Recipes)
- Poly-D-lysine stock solution (see Recipes)
- Treatment medium (see Recipes)
Equipment
- Cell culture incubator
- Centrifuges
- Dissection microscope
- Scissors and forceps
- T175 flasks
- 70 µm nylon filter (sterile)
- 50 ml tube
- Sterile filter (0.2 µm)
- Foil
- Laminar hood
Procedure
Category
Neuroscience > Cellular mechanisms > Cell isolation and culture
Cell Biology > Tissue analysis > Tissue isolation
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