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. 2018 Feb 1;29(1):44–59. doi: 10.1089/hgtb.2017.082

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© S. Louise Pay et al. 2018; Published by Mary Ann Liebert, Inc.

This article is available under the Creative Commons License CC-BY-NC (http://creativecommons.org/licenses/by-nc/4.0). This license permits non-commercial use, distribution and reproduction in any medium, provided the original work is properly cited. Permission only needs to be obtained for commercial use and can be done via RightsLink.

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Figure 5. — Efficacy of ILV3 and IDLV3 programmed BMDCs in a murine model of retinal degeneration. C57BL6/J mice received sub-retinal injection of AAV1-Rz-SOD2 or AAV1-Rz-inactive 1 month prior to tail-vein injections of naïve GFP⁺ murine BMDCs (null) or GFP⁺ BMDCs programmed with ILV3-RPE65 or IDLV3-RPE65. Animals were assessed 3 months following administration of BDMCs. (A) Visual acuity testing using an Optomotor behavioral test demonstrated recovery of optokinetic response in SOD2 knockdown mice receiving ILV3-RPE65-programmed BMDCs or IDLV3-RPE65-programmed BMDCs, which was not observed in animal receiving naïve BMDCs (null). A comparison of average values for photopic activities in response to rotating sinusoidal gratings is shown for each group of mice (n = 5 per group) as mean ± SEM. (B) Spectral domain optical coherence tomography (SD-OCT) was used to examine retinal morphology of the mouse retina in vivo. Major abnormalities were observed in the RPE/photoreceptor layer in RzSOD2 knockdown eyes with or without null BMDC treatment. However, the RzSOD2 knockdown eyes that received the ILV3-RPE65-programmed BMDC or IDLV3-RPE65-programmed BMDC treatments show near-normal retina architectures. (C) Representative fluorescent micrographs of flat-mounted RPE showing the presence of significant numbers of GFP⁺ (green) IDLV3-RPE65-BMDC and ILV3-RPE65-BMDCs. Only very few GFP⁺ cells were observed in mice treated systemically with null BMDCs. (D) Representative photomicrographs of 6 μM hematoxylin and eosin–stained cross-sections of the retina/choroid/sclera showed significant disruption of the RPE layer in the SOD2-KD mice, which was largely absent in mice receiving either ILV3-RPE65-programmed BMDC or IDLV3-RPE65-programmed BMDC, which both showed much improved retinal morphology. Higher magnification images of the RPE layer demonstrated that systemic administration of RPE65-programmed BMDCs with either ILV3 or IDLV3 have a near normal RPE morphology, similar to inactive Rz injected eyes. This was not apparent in animals receiving null BMDCs, in which there is thinning of the retina and severe loss of the RPE cells. RGC, retinal ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer; RPE, retinal pigment epithelium; CHO, choroid.