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Laura Liu, Yuntao Hu, Danhong Zhu, Padmaja B. Thomas, Kelsey Ford, Ashish K. Ahuja, Biju B. Thomas, Dennis O. Clegg, David R. Hinton, Mark S. Humayun; Human Embryonic Stem Cell Derived Retinal Pigment Epithelial Cells Cultured on Polymer Substrates Maintain a Confluent Monolayer Structure in the Subretinal Space of Implanted rats. Invest. Ophthalmol. Vis. Sci. 2011;52(14):3190.
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Human embryonic stem cell derived retinal pigment epithelial cells (hESC-RPE) can be cultured on the biocompatible polymer parylene as a monolayer for transplantation into the subretinal space. The purpose of this study is to evaluate the preservation of hESC-RPE seeded parylene substrates in implanted dystrophic and normal rats, and the ability of monolayer integrity to be preserved.
Polarized hESC-RPE cells (H9) were cultured on 10µm thick parylene sheets for 3 weeks. Rectangular pieces (0.4mm x 0.9mm) of parylene were seeded with RPE cells, which formed a confluent monolayer. These grafts were implanted into the subretinal space of dystrophic Royal College of Surgeons (RCS) rats (n=4) and normal Copenhagen rats (n=3). Prednisolone was administered for rats through drinking water (0.002mg/liter) for the entire period of study. Pre- and post-operative examinations were performed using fluorescence angiography, autofluorescene imaging, and spectral-domain optical coherence tomography. Animals were euthanized and eyes enucleated one month after surgery; tissue sections were subjected to histological evaluation using hemotoxylin and eosin staining.
Presence of hESC-RPE cells in the subretinal space was observed in all implanted rats. Architecture of confluent monolayer of RPE attached to the parylene was observed in the majority of rats (86%). No significant difference was observed between the RCS rats and the Copenhagen rats in terms of survival of transplanted cells. There were no obvious signs of ocular tumor formation or inflammation in sectioned samples up to one month after surgery.
hESC-RPE cultured on parylene substrates can be successfully implanted into the subretinal space of rats. Derived RPE cells can survive as a monolayer for at least one month without immune rejection when immunosuppressants are used. These results suggest such cell-seeded parylene implants can support hESC-RPE viability, and, therefore, be considered as a possible treatment for slowing the progression of dry-type age-related macular degeneration.
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