Cambridge Centre for Brain Repair, University of Cambridge, United Kingdom.
Invest Ophthalmol Vis Sci. 2011 May 17;52(6):3309-20. doi: 10.1167/iovs.10-6873.
PURPOSE. To validate an established adult organotypic retinal explant culture system for use as an efficient medium-throughput screening tool to investigate novel retinal ganglion cell (RGC) neuroprotective therapies. METHODS. Optimal culture conditions for detecting RGC neuroprotection in rat retinal explants were identified. Retinal explants were treated with various recognized, or purported, neuroprotective agents and cultured for either 4 or 7 days ex vivo. The number of cells surviving in the RGC layer (RGCL) was quantified using histologic and immunohistochemical techniques, and statistical analyses were applied to detect neuroprotective effects. RESULTS. The ability to replicate previously reported in vivo RGC neuroprotection in retinal explants was verified by demonstrating that caspase inhibition, brain-derived neurotrophic factor treatment, and stem cell transplantation all reduced RGCL cell loss in this model. Further screening of potential neuroprotective pharmacologic agents demonstrated that betaxolol, losartan, tafluprost, and simvastatin all alleviated RGCL cell loss in retinal explants, supporting previous reports. However, treatment with brimonidine did not protect RGCL neurons from death in retinal explant cultures. Explants cultured for 4 days ex vivo proved most sensitive for detecting neuroprotection. CONCLUSIONS. The current adult rat retinal explant culture model offers advantages over other models for screening potential neuroprotective drugs, including maintenance of neurons in situ, control of environmental conditions, and dissociation from other factors such as intraocular pressure. Verification that neuroprotection by previously identified RGC-protective therapies could be replicated in adult retinal explant cultures suggests that this model could be used for efficient medium-throughput screening of novel neuroprotective therapies for retinal neurodegenerative disease.
目的。验证已建立的成人器官型视网膜培养系统，以作为一种有效的高通量筛选工具，用于研究新型视网膜神经节细胞 (RGC) 神经保护疗法。
方法。确定了用于检测大鼠视网膜培养物中 RGC 神经保护作用的最佳培养条件。用各种公认的或据称的神经保护剂处理视网膜培养物，并在体外培养 4 或 7 天。用组织学和免疫组织化学技术定量检测 RGC 层 (RGCL) 中存活的细胞数量，并应用统计分析来检测神经保护作用。
结果。通过证明 caspase 抑制、脑源性神经营养因子治疗和干细胞移植均可减少该模型中 RGCL 细胞丢失，验证了在视网膜培养物中复制先前报道的体内 RGC 神经保护作用的能力。进一步筛选潜在的神经保护药物表明，倍他洛尔、洛沙坦、他氟前列素和辛伐他汀均可减轻视网膜培养物中 RGCL 细胞丢失，支持先前的报道。然而，在视网膜培养物中，用溴莫尼定治疗并不能保护 RGCL 神经元免于死亡。在体外培养 4 天的培养物最适合检测神经保护作用。
结论。目前的成年大鼠视网膜培养物模型在筛选潜在的神经保护药物方面具有优于其他模型的优势，包括原位维持神经元、控制环境条件以及与眼压等其他因素分离。先前确定的 RGC 保护疗法的神经保护作用可在成年视网膜培养物中复制，这表明该模型可用于对视网膜神经退行性疾病的新型神经保护疗法进行有效的高通量筛选。
