Related Experiment Video
Updated: Oct 3, 2025
Preparation of Primary Mixed Glial Cultures from Adult Mouse Spinal Cord Tissue
Published on: November 19, 2016
A new method for primary culture of microglia in rats with spinal cord injury
Chengjie Wu1, Yalan Pan2, Lining Wang3
1Department of Traumatology and Orthopedics, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, China; Laboratory of New Techniques of Restoration & Reconstruction, Institute of Traumatology & Orthopedics, Nanjing University of Chinese Medicine, Nanjing, China.
Abstract:
At present, the primary culture method of microglia is complicated, and the culture of spinal cord microglia is rare, so we will explore to establish a new and efficient primary culture method of microglia in rats with spinal cord injury (SCI). The SCI model of SD rats was established by modified A11en's method, and the model of SCI was performed on 1 d, 3 d, 7 d and 14 d respectively. Then the injured spinal cord was removed, mechanically separated and filtered. The morphology of microglia was observed the next day and its purity was identified by CD11b and Iba1 immunofluorescence labeling. According to the above results, the morphological changes of microglia after 3 d of SCI were observed at 1 d, 2 d and 4 d. The results showed that the purity of microglia was 98%. The number of microglia after 3 d of SCI was the most. After SCI, the migration ability of microglia was enhanced, the number of microglia in the injured area increased, and the number was the highest at 3 d, then gradually decreased. In addition, the microglia after SCI would gradually change from active state to resting state with the passage of time. Therefore, we can use a simple and efficient mechanical separation method to extract primary microglia, which provides the basis for the study of microglia.
Insights
Researchers developed a simple method to culture rat spinal cord microglia after injury. This efficient technique yields high-purity microglia, crucial for studying neuroinflammation and recovery post-spinal cord injury.
Area of Science:
- Neuroscience
- Cell Biology
- Immunology
Background:
- Primary microglia culture is complex, especially from injured spinal cords.
- Efficient methods are needed for studying microglia in spinal cord injury (SCI) models.
Purpose of the Study:
- To establish a novel, efficient primary culture method for microglia from rat spinal cords following SCI.
- To characterize microglia behavior and purity using this new method.
Main Methods:
- Established a rat spinal cord injury (SCI) model using a modified Allen's method.
- Utilized mechanical separation and filtration for microglia extraction.
- Assessed microglia morphology and purity via CD11b and Iba1 immunofluorescence labeling.
Main Results:
- Achieved 98% microglia purity using the mechanical separation method.
- Microglia numbers peaked at 3 days post-SCI, indicating enhanced migration.
- Observed microglia transitioning from an active to a resting state over time post-injury.
Conclusions:
- A simple, efficient mechanical separation technique effectively isolates primary microglia from injured rat spinal cords.
- This method provides a valuable foundation for future research on microglia in SCI.
- The findings offer insights into microglia dynamics and activation states following spinal cord trauma.
