June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Safety and Functionality of a Retinal Pigment Epithelium Monolayer Transplantation derived from Human Embryonic Stem Cells in Yucatan Mini-Pigs
Author Affiliations & Notes
  • Michael Koss
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Ramiro Ribeiro
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Rodrigo Brantfernandes
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Aldo Oregon-Miranda
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Padmaja Thomas
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Biju Thomas
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Danhong Zhu
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Gerald Chader
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • David Hinton
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Mark Humayun
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Footnotes
    Commercial Relationships Michael Koss, None; Ramiro Ribeiro, None; Rodrigo Brantfernandes, None; Aldo Oregon-Miranda, None; Padmaja Thomas, None; Biju Thomas, None; Danhong Zhu, None; Gerald Chader, None; David Hinton, RPT (I), RPT (P); Mark Humayun, Bausch & Lomb (F), Bausch & Lomb (C), Bausch & Lomb (P), Bausch & Lomb (R), Bausch & Lomb (S), Alcon (C), Alcon (R), Iridex (P), Iridex (R), Replenish (I), Replenish (C), Replenish (R), Replenish (S), Second Sight (F), Second Sight (I), Second Sight (C), Second Sight (P), Second Sight (R), Second Sight (S), Regenerative Patch Technologies (I), Regenerative Patch Technologies (C)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 313. doi:
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      Michael Koss, Ramiro Ribeiro, Rodrigo Brantfernandes, Aldo Oregon-Miranda, Padmaja Thomas, Biju Thomas, Danhong Zhu, Gerald Chader, David Hinton, Mark Humayun; Safety and Functionality of a Retinal Pigment Epithelium Monolayer Transplantation derived from Human Embryonic Stem Cells in Yucatan Mini-Pigs. Invest. Ophthalmol. Vis. Sci. 2013;54(15):313.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: To determine safety and functionality of a subretinal transplantation of a human embryonic stem cell (hESC) derived retinal pigment epithelium (hESC-RPE) monolayer seeded onto a sub-micron parylene-C membrane in Yucatan mini-pig eyes.

Methods: Animals were handled in accord with the ARVO Statement on Use of Animals in Ophthalmic/Visual Research. Four 2-months old female Yucatan mini pigs underwent a 25g vitrectomy with creation of a localized subretinal bleb. After a limited peripheral retinotomy and enlargement of the sclerotomy the transplant was placed using a designed inserter. Three animals received differentiated hESC-RPE polarized cells, which were attached onto the membranes and one the parylene membrane without the cells (sham). Immunosuppression included oral cyclosporine (100 mg/day for the first month, 200 mg/day for the last two months) and oral prednisone (100mg/day) during the 3 month follow up, which included in-vivo SD-OCT, fluorescein angiography (FA). Ex-vivo histological analyses included hematoxylin and eosin with immunohistochemical anti-RPE65 (RPE cell marker), rhodopsin and TRA-1-85 (human cell marker) antibody evaluation.

Results: All animals reached the 3 month follow-up with thorough in-vivo documentation in 3 of 4 animals. One pig could not be evaluated due to cataract. Subretinal parylene with and without hESC-RPE implantation was safe and did not cause deposit formation associated with inflammatory deposit formation or PVR development at the surgical site. FA demonstrated normal choroidal and retinal vascularization. The RPE cell monolayer was immunopositive for TRA-1-85 and RPE-65 in the hESC-RPE group without migration of the cells off the implant. Rhodopsin stain was positive indicating photoreceptor RPE functionality.

Conclusions: The hESC-RPE parylene transplantation represents a polarized cell replacement therapy on an artificial Bruchs’ Membrane-like substrate. The present technique was found to be surgically safe and demonstrated cell survival and functionality over three months. It thus may be a viable treatment for dry AMD.

Keywords: 412 age-related macular degeneration • 721 stem cells • 695 retinal degenerations: cell biology  
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