April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Photoreceptor-like Cells From Reprogramming Cultured Mammalian Rpe Cells
Author Affiliations & Notes
  • Run-Tao Yan
    Ophthalmology, Univ of Alabama at Birmingham, Birmingham, Alabama
  • Xiumei Li
    Ophthalmology, Univ of Alabama at Birmingham, Birmingham, Alabama
  • Shu-Zhen Wang
    Ophthalmology, Univ of Alabama at Birmingham, Birmingham, Alabama
  • Footnotes
    Commercial Relationships  Run-Tao Yan, None; Xiumei Li, None; Shu-Zhen Wang, None
  • Footnotes
    Support  EyeSight Foundation of Alabama Research Grant FY2008-09-134, NIH/NEI grant EY011640, and Research to Prevent Blindness
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 436. doi:
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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)

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Abstract

Purpose: : 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.

Methods: : 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.

Results: : 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.

Conclusions: : The results indicate that RPE→ photoreceptor reprogramming can occur in primary cell cultures derived from porcine and mouse RPE.

Keywords: photoreceptors • regeneration • retinal pigment epithelium 
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