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Francisco Rosa Stefanini, Michael J Koss, Paulo Falabella, Marcel Pfister, David R Hinton, Biju B Thomas, Padmaja Thomas, Dennis O Clegg, Mark S Humayun; Comparative analysis of retinal layers after subretinal stem cell implantation in Yucatan mini-pigs. Invest. Ophthalmol. Vis. Sci. 2014;55(13):3999. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To compare the thickness of the retinal layers in hematoxylin and eosin (H&E) stained histologic tissue sections after surgery for implantation of human embryonic stem cell-derived retinal pigment epithelium (hESC-RPE) monolayer in Yucatan mini-pigs.
Five Yucatan mini-pigs underwent vitrectomy with the creation of a localized subretinal BSS bleb before the implantation of hESC-RPE polarized cells cultured on parylene membranes into the subretinal space. One month after surgery, Aperio Scanscope landmark analysis from a standardized area of the retina was performed. It included the measurement of ganglion cells (GCL), nerve fiber (NFL), inner nuclear (INL), inner plexiform (IPL), outer nuclear (ONL), outer plexiform layer (OPL), photoreceptors (PR), and total retina thickness including the RPE (RT) in the anatomical site of the bleb (BS), the site of implant (IS) and the non-bleb retina (NBS). Three different areas per site were analyzed at 40x magnification from 3 H&E sections.
All animals reached the 1 month follow-up with thorough in vivo and ex vivo documentation. The implant region had an artifactual retinal detachment in all histologic sections, thus the PR and RT was not measureable. Considering the distance between the GCL/NFL to the ONL, the average overall thickness was as follows: BS (166.5 ± 12.7 μm), NBS (138.9 ± 9.9 μm), IS (147.1 ± 15.1 μm). IPL thickness was as follows: NBS (33.4 ± 6.8 μm) in BS (43.0 ± 7.3 μm) and IS (44.8 ± 7.6 μm). The ONL was as follows: IS (18.5 ± 5.2) ,BS (37.7 ± 11.1) and NBS (35.5 ± 3.6).
Overall there were minimal changes in the retina in the different sites after surgical implantation. The procedure of BSS infusion to create a subretinal pocket into which a hESC-RPE polarized cells cultured on parylene membranes is implanted was well tolerated. More studies are ongoing to further evaluation of this approach.
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