Neonatal Rat Primary Microglia: Isolation, Culturing, and Selected Applications

We provide a detailed description on the methods that is routinely used in our laboratory for the isolation and culturing of microglia, with emphasis on the steps which are deemed most critical for obtaining pure and healthy cultures

Mingwei Ni

2010

Scholarcy highlights

  • Neurons are the major cell type in the central nervous system, their unique electrical excitability distinguishing them from other cell types
  • Microglia are isolated from the mixed cell culture before the end of the 2nd week by gentle physical shaking and tapping on the flasks
  • The first step focuses on dissecting rat cortices and the second one entails the dissociation of glial cells including astrocytes, microglia and small amount of oligodendrocytes
  • It is important to change culture media twice a week to keep cells healthy; otherwise, astrocytes could be sheared during the isolation procedure very
  • The primary microglia seeded and grown on Poly-L-lysine coated plastic surface do not grow in clusters, so that fine structures on the cell membrane could be observed in detail
  • The total time spent on the preparation of mixed glial culture is approximately 6 hours depending on the number of extractions performed and the number of rat pups used
  • Castellano B, Gonzalez B, et al A double staining technique for simultaneous demonstration of astrocytes and microglia in brain sections and astroglial cell cultures

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