Purchase this article with an account.
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.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
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.
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.
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.
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.
This PDF is available to Subscribers Only