Preparation of Primary Mixed Glial Cell Cultures from Adult Mouse Spinal Cord Tissue

Central nervous system glial cells play a role in neurocognitive and neurodegenerative diseases like Alzheimer’s and Parkinson’s. Similar glial responses are important in peripheral nervous system diseases, such as neuropathic pain and diabetic neuropathy. This paper describes a method for preparing mixed glial cells from adult mouse spinal cord for in vitro study. The described protocol involves enzymatically dissociating spinal cord tissue. The adult mixed glial cells are ready for use 12-14 days after culture establishment. This method can be used on spinal cord tissues from adult rats and potentially other species. Mixed glial cultures can be prepared from different animal strains or after in vivo manipulations. They are suitable for studying diseases involving spinal cord pathological changes like amyotrophic lateral sclerosis and multiple sclerosis, as well as toxin-induced changes.

• 1
  This protocol provides a method for preparing mixed glial cells from adult mouse spinal cord, suitable for in vitro evaluations or further preparation of microglia-enriched and microglia-depleted cells.
• 2
  The method is flexible and can be adapted to prepare primary mixed glial cells from rat spinal cord tissue and potentially other species.
• 3
  Mixed glial cultures can be prepared from various animal strains or in vivo-manipulated animals, enabling the study of diseases/disorders involving pathological changes within the spinal cord.