North Texas Eye Research Institute, Department of Cell Biology and Anatomy, University of North Texas Health Science Center, Fort Worth, Texas 76107, USA.
Invest Ophthalmol Vis Sci. 2013 Aug 23;54(8):5712-9. doi: 10.1167/iovs.13-12085.
In 2001, a transformed cell line RGC-5 was developed from the rat retina that was thought to be of retinal ganglion cell origin. Since that time many investigators have used this line in a wide variety of studies to understand better retinal ganglion cell activity, cell signaling, and neuroprotection. Recently, a publication emerged that claimed that this RGC-5 cell line was derived from mouse and not rat, and other studies also indicated the expression of certain proteins that typically were not associated with retinal ganglion cells. This certainly came as a shock not only to the originators of this cell line, but also to others who have been using this as an in vitro model of rat retinal ganglion cells. As a result, we undertook experiments to determine if the RGC-5 cell line currently in use may have been mischaracterized. We, indeed, found that the RGC-5 cell line was of mouse and not rat origin, as was claimed originally in the original research report. We further determined whether these cells were of retinal ganglion origin. Our findings showed conclusively that RGC-5 cells were, indeed, of mouse origin and, using additional cytogenetic profile testing, karyotyping, and genetic and protein profiling, we concluded that these cells were not of retinal ganglion cell origin, but were the cell line 661W, a mouse SV-40 T antigen transformed photoreceptor cell line. The 661W cell line also was present in the laboratory of the originating laboratory and probably resulted in cross-contamination. The present study reviews some of the errors that were made in misidentifying the RGC-5 cell line and offers some insight as to how this may have happened, and ways one can avoid mischaracterization of a potentially important cell line.
2001 年，从大鼠视网膜中开发出一种转化细胞系 RGC-5，该细胞系被认为来源于视网膜神经节细胞。自那时以来，许多研究人员在广泛的研究中使用该细胞系来更好地理解视网膜神经节细胞的活性、细胞信号转导和神经保护。最近，有一篇出版物声称该 RGC-5 细胞系源自小鼠而不是大鼠，其他研究也表明存在某些通常与视网膜神经节细胞无关的蛋白质的表达。这不仅令该细胞系的创建者感到震惊，也令其他一直将其作为大鼠视网膜神经节细胞体外模型使用的人感到震惊。因此，我们进行了实验以确定目前正在使用的 RGC-5 细胞系是否可能被错误描述。我们确实发现，正如原始研究报告中最初声称的那样，RGC-5 细胞系源自小鼠而不是大鼠。我们进一步确定这些细胞是否源自视网膜神经节。我们的研究结果明确表明，RGC-5 细胞确实源自小鼠，并且使用额外的细胞遗传学特征测试、核型分析、基因和蛋白质分析，我们得出结论，这些细胞不是源自视网膜神经节细胞，而是源自细胞系 661W，这是一种小鼠 SV-40 T 抗原转化的光感受器细胞系。661W 细胞系也存在于原始实验室的实验室中，可能导致了交叉污染。本研究回顾了在错误鉴定 RGC-5 细胞系时所犯的一些错误，并提供了一些有关这些错误如何发生的见解，以及如何避免对一种潜在重要的细胞系进行错误描述。
