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Padmaja B. Thomas, Biju B. Thomas, Laura Liu, Yuntao Hu, Danhong Zhu, Ernesto Barron, Dennis O. Clegg, David R. Hinton, Mark S. Humayun; Evaluation of hESC-Derived Retinal Pigment Epithelial Cells Cultured as a Monolayer on Polymer Substrate Transplanted in RCS Rats. Invest. Ophthalmol. Vis. Sci. 2012;53(14):5920. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate the survival and functionality of human embryonic stem cell (hESC)-derived retinal pigment epithelial (RPE) cells cultured on the biocompatible polymer parylene as a monolayer and transplanted into the subretinal space of Royal College of Surgeon (RCS) rats using histological techniques.
Polarized hESC-RPE cells (H9) were cultured for 4 weeks on parylene which formed a confluent monolayer and were implanted into the subretinal space of RCS rats. Prednisolone was administered to rats through drinking water (0.002mg/liter) for the entire period of study. Animals were euthanized and eyes enucleated 8 weeks post-transplantation. Tissue sections were subjected to histological evaluation based on Hemotoxylin and Eosin (H&E)and immunostaining. Cell counting was performed using an Apereo scanscope. Electron microscopy and immunostaining techniques were used to evaluate phagocytosis by hESC-RPE.
The presence of a confluent monolayer of hESC-RPE attached to the parylene was observed based on H&E staining. Survival of the implanted cells was confirmed by the expression of TRA-1-85 a human marker and RPE-65 which is an RPE marker. The transplanted RPE-cells were able to phagocytose host photoreceptor outer segments (POS). Based on cell counting, the number of photoreceptor nuclei preserved over the implanted area was significantly higher compared to the non-implanted area (p<0.05).
hESC-RPE cultured on parylene substrates can survive as a monolayer in the subretinal space of RCS rats for at least two months without immune rejection and were able to phagocytise host POS. hESC-RPE cell transplantation attenuates the progression of photoreceptor loss in the implanted eyes. These results suggest such cell-seeded parylene implants can support hESC-RPE survival and functionality, and therefore, be considered as a possible treatment for dry age-related macular degeneration.
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