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Run-Tao Yan, Xiumei Li, Shu-Zhen Wang; Photoreceptor-like Cells From Reprogramming Cultured Mammalian Rpe Cells. Invest. Ophthalmol. Vis. Sci. 2011;52(14):436.
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© ARVO (1962-2015); The Authors (2016-present)
Previous studies using the chick system provide compelling evidence for gene-directed reprogramming of RPE progeny cells to produce differentiating photoreceptors. In this study, we tested whether mammalian RPE cells can be guided by a pro-photoreceptor gene towards differentiating as photoreceptor cells.
Primary cell cultures were established with isolated RPE from porcine and mouse eyes. The culture was infected with a lenti virus (Lvx-IRES-ZsGreen) expressing human neurogenin1 (Lvx-ngn1-IRES-ZsGreen) or neurogenin3 (Lenti-virus Lvx-ngn3-IRES-ZsGreen1). Expression of the transgene was indirectly monitored by viewing ZsGreen1. Reprogramming of the transduced cells as differentiating photoreceptor cells were evaluated by morphology and immunostaining for photoreceptor proteins.
Three days after viral administration, some ZsGreen1+ cells, particularly those with brighter fluorescence (likely to have higher levels of transgene expression), began to exhibit morphologies resembling differentiating photoreceptor cells. Immunochemistry showed that some ZsGreen1+ cells were positive for neural maker Map2, rhodopsin, or red opsin. Photoreceptor-like cells were detected in both P5 mouse and adult porcine RPE cell cultures.
The results indicate that RPE→ photoreceptor reprogramming can occur in primary cell cultures derived from porcine and mouse RPE.
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