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Updated: May 30, 2026
Obtaining Human Microglia from Adult Human Brain Tissue
Published on: August 30, 2020
A much convenient and economical method to harvest a great number of microglia
1Neurosurgery Institute, Key Laboratory on Brain Function Repair and Regeneration of Guangdong, Zhujiang Hospital, Southern Medical University, 510282, Guangzhou, China.
Abstract:
Microglia, implicating in such neuro-pathologies as brain inflammation, neurodegeneration, glioma, and neurogenesis, play an important role in central nervous system. Advanced research on microglia is crucial in exploring the neuro-pathology and neuro-physiology of these diseases, so how to culture large numbers of microglia in vitro becomes the base of a research. The wildly used method, at present, obtaining microglia from murine cannot fulfill the requirement of research, costing too much time and needing too many rats. We intend to introduce an optimized method that can harvest large quantities of microglia with high purity. Neonatal 2-3 days old Wistar rats were sacrificed and the cerebral cortices were trypsinized. We primarily cultured mixed cortical cells for 8-10 days. The microglia were harvested from the liquid supernatant; the left cells in the mixed cortical glial culture were passaged at a 1:2 density. After another 8-10 days of culture, microglia were collected again. And then, we passaged the left cells again for acquiring microglia from the third collection. We did not add additional mitogens in the experiment. At last, on average, 7.0 × 10(6) microglia were collected from one neonatal rat. By this modified method, much more microglia can be effectively and easily harvested comparing with the usual protocol before. We compared the characteristics of microglia harvested from these three passages, such as morphology, phenotype, purity, and abilities on proliferation, secretion, and phagocytosis. The cells presented typical microglia morphology, having phenotype markers of CD11b/c and CD45. The microglia from these three passages retained similar phagocytosis and secretion functions. Expanded population of microglia for investigation can be provided by this easy method in a short time with little cost and few rats.
Insights
This study introduces an optimized method for culturing microglia, essential immune cells in the central nervous system. The new technique yields significantly more high-purity microglia from fewer rats, reducing research costs and time.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are crucial for central nervous system health and disease, including neuroinflammation and neurodegeneration.
- Current methods for obtaining microglia in vitro are time-consuming and require a large number of animals.
- Efficient in vitro microglia culture is essential for advancing research in neuro-pathology and neuro-physiology.
Purpose of the Study:
- To develop and validate an optimized method for harvesting large quantities of high-purity microglia from neonatal rats.
- To compare the yield and characteristics of microglia obtained through the modified protocol versus traditional methods.
- To provide a cost-effective and efficient approach for microglia expansion for research purposes.
Main Methods:
- Neonatal Wistar rat cerebral cortices were trypsinized and primary mixed cortical cells were cultured.
- Microglia were harvested from the supernatant after initial culturing and subsequent passaging.
- Cells were passaged twice more, with microglia collected after each 8-10 day culture period, without additional mitogens.
Main Results:
- The optimized method yielded an average of 7.0 × 10^6 microglia per neonatal rat.
- Microglia from three passages exhibited typical morphology and expressed CD11b/c and CD45 phenotype markers.
- All three passages of microglia demonstrated similar phagocytosis and secretion functions.
Conclusions:
- This modified protocol significantly increases microglia yield and purity compared to conventional methods.
- The method is efficient, cost-effective, and requires fewer animals, making it suitable for large-scale microglia expansion.
- The harvested microglia maintain their characteristic functions, providing a valuable resource for neuroscience research.
