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G.M. Zainelli, B.C. Barron, J. Wen, J.A. Kapp, E. Garcia; Implantation of neural retinal precursor cells (NRPCs) into the subretinal space of light–damaged mouse eyes. . Invest. Ophthalmol. Vis. Sci. 2004;45(13):5411.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Repair light–induced retinal damage in C57/BL6 and BALB/c mice by subretinal injection of neural retinal precursor cells (NRPCs) Methods: C57/BL6 and BALB/c mice (4–6 weeks old) were dark–adapted for 24h. Their pupils were dilated and they were exposed to 20–22,000 lux of cool, white light for up to 8h, in a white–reflective chamber with controlled, ambient temperature (27ºC). Ten days later, mice were sacrificed and their eyes were fixed in either 4% paraformaldehyde for frozen sections, or formalin for paraffin sections. Frozen sections were stained with either DAPI or propidium iodide to visualize nuclei. Paraffin–embedded sections (5µM) were then stained with Hematoxylin and Eosin (H&E stain). NRPCs were isolated from the ciliary margin of C57/BL6 mice and cultured in a low serum media containing 1% fetal bovine serum (FBS) for 5–7 days. NRPCs were then implanted into the subretinal space of eyes of mice 7–10 days after the phototoxic insult. Loss of cells in the outer nuclear layer of the retina, and integration of NRPCs was assessed using immunohistochemistry. Results: Very sparse inner and outer nuclear layer cell death was observed in C57/BL6 mice that were exposed to 6h of the phototoxic insult. In contrast, high levels of outer nuclear layer and photoreceptor layer death were observed in BALB/c mice after as little as 2h of the phototoxic insult. Implantation of NRPCs to the subretinal space did not cause an inflammatory response in C57/BL6 mice when examined at ten days after transplantation. The survival of NRPCs and the ability of NRPCs to integrate into the retina are currently under investigation. Conclusions:Histological analysis of tissue sections following phototoxic insult and subsequent subretinal injections of NRPCs constitute a model system for stem cell transplantation and integration into the damaged or diseased retina.
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