Center for Ophthalmology, University Eye Hospital Tübingen, Elfriede-Aulhorn-Str. 7, 72076 Tuebingen, Germany.
Institute for Medical Genetics and Applied Genomics, Center for Rare Diseases, University of Tuebingen, Calwerstrasse 7, 72076 Tübingen, Germany.
Int J Mol Sci. 2023 Sep 7;24(18):13801. doi: 10.3390/ijms241813801.
In the pursuit of identifying the underlying pathways of ocular diseases, the use of cell lines such as (retinal ganglion cell-5) RGC-5 and 661W became a valuable tool, including pathologies like retinal degeneration and glaucoma. In 2001, the establishment of the RGC-5 cell line marked a significant breakthrough in glaucoma research. Over time, however, concerns arose about the true nature of RGC-5 cells, with conflicting findings in the literature regarding their identity as retinal ganglion cells or photoreceptor-like cells. This study aimed to address the controversy surrounding the RGC-5 cell line's origin and properties by comparing it with the 661W cell line, a known cone photoreceptor model. Both cell lines were differentiated according to two prior published redifferentiation protocols under the same conditions using 500 nM of trichostatin A (TSA) and investigated for their morphological and neuronal marker properties. The results demonstrated that both cell lines are murine, and they exhibited distinct morphological and neuronal marker properties. Notably, the RGC-5 cells showed higher expression of the neuronal marker β-III tubulin and increased Thy-1-mRNA compared with the 661W cells, providing evidence of their different properties. The findings emphasize the importance of verifying the authenticity of cell lines used in ocular research and highlight the risks of contamination and altered cell properties.
在探索眼部疾病的潜在发病机制的过程中，（视网膜神经节细胞-5）RGC-5 和 661W 等细胞系的应用成为了一种有价值的工具，可用于研究视网膜变性和青光眼等疾病。2001 年，RGC-5 细胞系的建立标志着青光眼研究的重大突破。然而，随着时间的推移，人们对 RGC-5 细胞的真实性质产生了担忧，文献中关于其作为视网膜神经节细胞或光感受器样细胞的身份存在相互矛盾的发现。本研究旨在通过与已知的视锥细胞模型 661W 细胞系进行比较，来解决 RGC-5 细胞系起源和特性的争议。这两种细胞系均根据之前发表的两种再分化方案，在相同条件下使用 500 nM 的曲古抑菌素 A（TSA）进行分化，并对其形态和神经元标记特性进行了研究。结果表明，这两种细胞系均为鼠源性细胞，具有明显不同的形态和神经元标记特性。值得注意的是，与 661W 细胞相比，RGC-5 细胞β-III 微管蛋白的表达更高，Thy-1-mRNA 的表达也增加，这为它们的不同特性提供了证据。这些发现强调了在眼部研究中验证所使用细胞系真实性的重要性，并突出了污染和细胞特性改变的风险。
