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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 \u00b1 4% and 67.6 \u00b1 3% in control and COH groups, respectively. The proportion of the microglia population in the whole retinal cells was about 0.4\u20130.93%. 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.<\/jats:p>","DOI":"10.1515\/biol-2021-0100","type":"journal-article","created":{"date-parts":[[2021,9,15]],"date-time":"2021-09-15T20:53:45Z","timestamp":1631739225000},"page":"992-1001","source":"Crossref","is-referenced-by-count":6,"title":["Isolation of microglia from retinas of chronic ocular hypertensive rats"],"prefix":"10.1515","volume":"16","author":[{"given":"Huimin","family":"Zhong","sequence":"first","affiliation":[{"name":"Department of Ophthalmology, Ruijin Hospital Affiliated Medical School, Shanghai Jiaotong University , 197 Ruijin Er Road , 200025 , Shanghai , China"},{"name":"Department of Ophthalmology, Shanghai General Hospital (Shanghai First People s Hospital), National Clinical Research Center for Eye Diseases, Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases , Shanghai , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huan","family":"Yu","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Ruijin Hospital Affiliated Medical School, Shanghai Jiaotong University , 197 Ruijin Er Road , 200025 , Shanghai , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun","family":"Sun","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Ruijin Hospital Affiliated Medical School, Shanghai Jiaotong University , 197 Ruijin Er Road , 200025 , Shanghai , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Junjue","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Ruijin Hospital Affiliated Medical School, Shanghai Jiaotong University , 197 Ruijin Er Road , 200025 , Shanghai , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shouyue","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Ruijin Hospital Affiliated Medical School, Shanghai Jiaotong University , 197 Ruijin Er Road , 200025 , Shanghai , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ping","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Orthopaedics, Shanghai Key Laboratory for Prevention and Treatment of Bone and Joint Diseases, Shanghai Institute of Traumatology and Orthopaedics, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine , 197 Ruijin 2nd Road , Shanghai 200025 , People\u2019s Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaohong","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Ruijin Hospital Affiliated Medical School, Shanghai Jiaotong University , 197 Ruijin Er Road , 200025 , Shanghai , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yisheng","family":"Zhong","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Ruijin Hospital Affiliated Medical School, Shanghai Jiaotong University , 197 Ruijin Er Road , 200025 , Shanghai , China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"374","published-online":{"date-parts":[[2021,9,15]]},"reference":[{"key":"2022092006444916049_j_biol-2021-0100_ref_001","doi-asserted-by":"crossref","unstructured":"Zhang Y, Erhard AL, Plagemann T, Eter N, Heiduschka P. 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