April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Comparative analysis of retinal layers after subretinal stem cell implantation in Yucatan mini-pigs
Author Affiliations & Notes
  • Francisco Rosa Stefanini
    Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, Federal University of Sao Paulo, Sao Paulo, Brazil
  • Michael J Koss
    Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, University of Southern California, Los Angeles, CA
  • Paulo Falabella
    Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, Federal University of Sao Paulo, Sao Paulo, Brazil
  • Marcel Pfister
    Doheny Eye Institute, Los Angeles, CA
  • David R Hinton
    Ophthalmology, University of Southern California, Los Angeles, CA
  • Biju B Thomas
    Ophthalmology, University of Southern California, Los Angeles, CA
  • Padmaja Thomas
    Doheny Eye Institute, Los Angeles, CA
  • Dennis O Clegg
    Univ of California-Santa Barbara, Santa Barbara, CA
  • Mark S Humayun
    Ophthalmology, University of Southern California, Los Angeles, CA
  • Footnotes
    Commercial Relationships Francisco Stefanini, None; Michael Koss, None; Paulo Falabella, None; Marcel Pfister, None; David Hinton, None; Biju Thomas, None; Padmaja Thomas, None; Dennis Clegg, None; Mark Humayun, Baush & Lomb (C)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 3999. doi:https://doi.org/
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    • Get Citation

      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)

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Abstract

Purpose: 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.

Methods: 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.

Results: 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).

Conclusions: 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.

Keywords: 721 stem cells • 701 retinal pigment epithelium • 688 retina  
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