Isolation of microglia from retinas of chronic ocular hypertensive rats
| Autor: | Shouyue Huang, Jun Sun, Ping Huang, Yisheng Zhong, Xiaohong Liu, Huan Yu, Huimin Zhong, Junjue Chen |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Pathology
medicine.medical_specialty QH301-705.5 Population Context (language use) Biology General Biochemistry Genetics and Molecular Biology retinal microglia chemistry.chemical_compound medicine Biology (General) education education.field_of_study Retina General Immunology and Microbiology Microglia General Neuroscience Retinal eye diseases primary cells separation medicine.anatomical_structure nervous system chemistry Integrin alpha M coh biology.protein Optic nerve sense organs percoll gradient General Agricultural and Biological Sciences Percoll Research Article |
| Zdroj: | Open Life Sciences, Vol 16, Iss 1, Pp 992-1001 (2021) Open Life Sciences |
| ISSN: | 2391-5412 |
| Popis: | Microglia are the principal glial cells involved in the processes of immune inflammation within both retina and optic nerve, especially under the context of glaucomatous neuropathy. Considering the distinguishing role of retinal microglia in glaucoma and the lack of established protocol for microglia isolation from animal glaucoma model, the present study aimed to develop and validate a method with characteristics of both simplicity and efficiency for retinal microglia isolation from chronic ocular hypertensive (COH) rats. A Percoll gradient of various concentrations was used to separate microglia from whole retinal cells of the COH rats and control group. The finally isolated microglia were identified by CD11b and Iba-1 immunofluorescence staining, and the cell viability was determined by trypan blue staining. Additionally, the proportion of microglia in the whole retina cells was identified by flow cytometry. Results showed that the survival rates of isolated retinal microglia with the Percoll gradient method were 67.2 ± 4% and 67.6 ± 3% in control and COH groups, respectively. The proportion of the microglia population in the whole retinal cells was about 0.4–0.93%. To conclude, the present study confirmed that the application of Percoll gradient could effectively separate microglia from retinas of COH rats, which will probably enrich the tool kit for basic researchers of glaucoma specialty and help with scientific investigations. Graphical abstract The application of Percoll gradient method could effectively separate microglia cells from retinas of COH rats. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|