March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Evaluation of hESC-Derived Retinal Pigment Epithelial Cells Cultured as a Monolayer on Polymer Substrate Transplanted in RCS Rats
Author Affiliations & Notes
  • Padmaja B. Thomas
    Doheny Eye Institute-USC, Los Angeles, California
  • Biju B. Thomas
    Ophthalmology,
    Doheny Eye Institute-USC, Los Angeles, California
  • Laura Liu
    Cell and Neurobiology, University of Southern Califormia, Los Angeles, California
    Ophthalmology, Chang Gung Memorial Hospital, Taoyuan, Taiwan
  • Yuntao Hu
    Doheny Eye Institute-USC, Los Angeles, California
    Ophthalmology, Peking University Third Hospital, Beijing, China
  • Danhong Zhu
    Pathology/Doheny Eye Inst, Univ of Southern California, Los Angeles, California
  • Ernesto Barron
    Doheny Eye Institute-USC, Los Angeles, California
  • Dennis O. Clegg
    Bioscience II, Center for Stem Cell Biology and Engineering- UCSB, Santa Barbara, California
  • David R. Hinton
    Pathology, Keck School of Medicine USC, Los Angeles, California
  • Mark S. Humayun
    Ophthalmology, Doheny Eye Institute - USC, Los Angeles, California
  • Footnotes
    Commercial Relationships  Padmaja B. Thomas, None; Biju B. Thomas, None; Laura Liu, None; Yuntao Hu, None; Danhong Zhu, None; Ernesto Barron, None; Dennis O. Clegg, None; David R. Hinton, None; Mark S. Humayun, None
  • Footnotes
    Support  California Institute for Regenerative Medicine (CIRM), NEI EY03040
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 5920. doi:https://doi.org/
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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)

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Abstract

Purpose: : 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.

Methods: : 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.

Results: : 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).

Conclusions: : 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.

Keywords: retinal pigment epithelium • transplantation • age-related macular degeneration 
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