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Primary Microglia Isolation from Postnatal Mouse Brains
Siling Du1, Shanshan Xiong2, Xiangjuan Du3
1Shanghai Key Laboratory of Psychotic Disorders, Shanghai Mental Health Center, Shanghai Jiao Tong University School of Medicine; Division of Biology and Biomedical Science, Washington University in St. Louis.
Abstract:
Microglia are the mononuclear phagocytes in the central nervous system (CNS), which play key roles in maintaining homeostasis and regulating the inflammatory process in the CNS. To study the microglial biology in vitro, primary microglia show great advantages compared to immortalized microglial cell lines. However, microglia isolation from the postnatal mouse brain is relatively less efficient and time-consuming. In this protocol, we provide a quick and easy-to-follow method to isolate primary microglia from the neonatal mouse brain. The overall steps of this protocol include brain dissection, primary brain cell culture, and microglia isolation. Using this approach, researchers can obtain primary microglia with high purity. In addition, the harvested primary microglia were able to respond to the lipopolysaccharides challenge, indicating they retained their immune function. Collectively, we developed a simplified approach to efficiently isolate primary microglia with high purity, which facilitates a wide range of microglial biology investigations in vitro.
Insights
This study presents a simplified protocol for efficiently isolating primary microglia from neonatal mouse brains. The method yields high-purity microglia, crucial for studying central nervous system inflammation and homeostasis.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are essential for central nervous system (CNS) homeostasis and inflammation.
- Primary microglia are superior to cell lines for in vitro studies of microglial biology.
- Existing methods for isolating primary microglia are inefficient and time-consuming.
Purpose of the Study:
- To develop a quick, easy, and efficient protocol for isolating primary microglia from neonatal mouse brains.
- To provide researchers with a reliable method for obtaining high-purity primary microglia for in vitro investigations.
Main Methods:
- The protocol involves brain dissection, primary brain cell culture, and microglia isolation.
- This method is designed to be straightforward and accessible for researchers.
Main Results:
- The developed protocol successfully isolates primary microglia with high purity.
- Isolated primary microglia demonstrate retained immune function, responding to lipopolysaccharide challenges.
Conclusions:
- A simplified and efficient method for isolating primary microglia has been established.
- This protocol facilitates extensive in vitro investigations into microglial biology and CNS inflammatory processes.
