Isolation of microglia from aged rat brain tissue and phenotypic identification by flow cytometry
10.19405/j.cnki.issn1000-1492.2024.11.011
- Author:
Miaomiao Liu
1
;
Meiqi Guan
2
;
Lian Yu
1
;
Xiaorong Yang
3
;
Jie Wang
1
;
Yanli Li
1
Author Information
1. Dept of Neurology , The First Hospital of Shanxi Medical University , Taiyuan 030001
2. Dept of Pediatrics , Ministry of Education , Dept of Physiology , Shanxi Medical University , Taiyuan 030001
3. Key Laboratory of Cellular Physiology , Ministry of Education , Dept of Physiology , Shanxi Medical University , Taiyuan 030001
- Publication Type:Journal Article
- Keywords:
aging;
neuroinflammation;
isolation and purification;
microglia;
flow cytometry;
surface marker
- From:
Acta Universitatis Medicinalis Anhui
2024;59(11):1967-1973
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish an optimized method for isolation and purification of microglia from aged rat brain tissue, and the phenotype of microglia was detected by flow cytometry.
Methods:With young rats(3 months old) as control, the brain tissues of aged rats were immediately processed into single cell suspensions by mechanical dissociation and enzymatic digestion using type IV collagenase. Microglia were isolated on Percoll gradients(30%-37%-70%). The cells were stained with fluorescence-labeled antibodies and the phenotype of microglia was detected by flow cytometry.
Results:This study developed a method that enzymatic digestion and mechanical dissociation combined with density gradient centrifugation. More single cells could be obtained by using this method. And the survival rate of cells was more than 90%. The flow cytometric analysis showed that the expression of M1 microglia marker CD86 and MHC Ⅱ increased(P<0.01), and the expression of M2 microglia marker CD200R increased(P<0.01) in aged rats compared with that in young rats.
Conclusion:The use of type IV collagenase and mechanical digestion combined with density gradient centrifugation is good for isolating and purifying microglia from adult and aged rat brain tissue.
- Full text:2024122613260492229老年大鼠脑组织小胶质细胞的分离及表型的流式鉴定_刘苗苗.pdf
