title: Viability of primary cultured retinal neurons in a hyperglycemic condition.
abstract: The retina of Wistar rats within 1-3 days of birth were dissociated into a retinal cell suspension using 0.05% trypsin digestion. The cell suspension was incubated in Dulbecco's modified Eagle's medium for 24 hours, followed by neurobasal medium for 5-7 days. Nissl staining showed that 79.86% of primary cultured retinal cells were positive and immunocytochemical staining showed that the purity of anti-neurofilament heavy chain antibody-positive cells was 71.53%, indicating that the primary culture system of rat retinal neurons was a reliable and stable cell system with neurons as the predominant cell type. The primary cultured retinal neurons were further treated with 0, 5.5, 15, 25, and 35 mM glucose for 24, 48, and 72 hours. The thiazolyl blue tetrazolium bromide test and flow cytometry showed that with increasing glucose concentration and treatment duration, the viability of retinal neurons was reduced, and apoptosis increased. In particular, 35 mM glucose exhibited the most significant effect at 72 hours. Thus, rat retinal neurons treated with 35 mM glucose for 72 hours can be used to simulate a neuronal model of diabetic retinopathy.
authors.affiliation: Department of Ophthalmology, Third Xiangya Hospital of Central South University, Changsha 410013, Hunan Province, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Ophthalmology, Xiangya Hospital of Central South University, Changsha 410008, Hunan Province, China.
authors.forename: Xueliang
authors.initials: X
authors.lastname: Xu
authors.affiliation: Department of Ophthalmology, Third Xiangya Hospital of Central South University, Changsha 410013, Hunan Province, China.
authors.forename: Renhong
authors.initials: R
authors.lastname: Tang
authors.affiliation: Department of Ophthalmology, Third Xiangya Hospital of Central South University, Changsha 410013, Hunan Province, China.
authors.forename: Guoping
authors.initials: G
authors.lastname: Chen
authors.affiliation: Department of Ophthalmology, Third Xiangya Hospital of Central South University, Changsha 410013, Hunan Province, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Lei
authors.affiliation: Department of Ophthalmology, Third Xiangya Hospital of Central South University, Changsha 410013, Hunan Province, China.
authors.forename: Limo
authors.initials: L
authors.lastname: Gao
authors.affiliation: Department of Ophthalmology, Third Xiangya Hospital of Central South University, Changsha 410013, Hunan Province, China.
authors.forename: Wenjie
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Ophthalmology, Third Xiangya Hospital of Central South University, Changsha 410013, Hunan Province, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Chen
keywords: apoptosis; diabetic retinopathy; glucose; grants-supported paper; hyperglycemia model; neural regeneration; neurons; neuroregeneration; peripheral nerve injury; photographs-containing paper; retina