March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Behavioral and Electrophysiological Evaluation of Long-term Functionality of hESC Derived RPE Monolayer Implanted in RCS Rats
Author Affiliations & Notes
  • Biju B. Thomas
    Ophthalmology,
    Doheny Eye Institute - USC, Los Angeles, California
  • Alejandra Gonzalez-Calle
    Doheny Eye Institute - USC, Los Angeles, California
  • Yuntao Hu
    Ophthalmology,
    Doheny Eye Institute - USC, Los Angeles, California
    Ophthalmology, Peking University Third Hospital, Beijing, China
  • Laura Liu
    Cell and Neurobiology, University of Southern California, Los Angeles, California
    Ophthalmology, Chang Gung Memorial Hospital, Taoyuan, Taiwan, Taiwan
  • Danhong Zhu
    Pathology/Doheny Eye Inst, Univ of Southern California, Los Angeles, California
  • Padmaja B. Thomas
    Doheny Eye Institute - USC, Los Angeles, California
  • Ann L. Philip
    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  Biju B. Thomas, None; Alejandra Gonzalez-Calle, None; Yuntao Hu, None; Laura Liu, None; Danhong Zhu, None; Padmaja B. Thomas, None; Ann L. Philip, 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, 313. doi:https://doi.org/
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      Biju B. Thomas, Alejandra Gonzalez-Calle, Yuntao Hu, Laura Liu, Danhong Zhu, Padmaja B. Thomas, Ann L. Philip, Dennis O. Clegg, David R. Hinton, Mark S. Humayun; Behavioral and Electrophysiological Evaluation of Long-term Functionality of hESC Derived RPE Monolayer Implanted in RCS Rats. Invest. Ophthalmol. Vis. Sci. 2012;53(14):313. doi: https://doi.org/.

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

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Abstract

Purpose: : Long-term evaluation of human embryonic stem cell (hESC) derived retinal pigment epithelial (RPE) cells cultured on the biocompatible polymer parylene as a monolayer transplanted into the subretinal space of Royal College of Surgeon (RCS) rats using optokinetic testing and superior colliculus (SC) electrophysiology.

Methods: : Polarized hESC-RPE cells (H9) were cultured for 4 weeks on parylene sheets. 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 RCS rats (n=6). Rats that received parylene sheets alone (without hESC-RPE) served as the control group (n=3). All animals were administered with oral cyclosporine A (CyA in drinking water, 210mg/l) from one day prior to surgery and maintained for the entire period of study. An intraperitoneal injection of dexamethasone was given for the initial two weeks (1.6mg/kg/day) after surgery. Post-operative examinations were performed using spectral-domain optical coherence tomography (OCT) and optokinetic testing. Electrophysiological mapping of the SC was performed at 6 months post-implantation.

Results: : OCT examination confirmed the placement of the hESC-RPE graft in the rat’s subretinal space. Optokinetic testing demonstrated considerably higher visual activity only in the hESC-RPE transplanted eye and was maintained throughout the study period. Electrophysiological testing revealed that visual responses recorded from transplanted rats under low level light stimulation (threshold = -1.0 log cd/m2) was confined to a small area on the surface of the SC. Fundus imaging data confirmed that the location of the hESC-RPE graft inside the eye is corresponding to the area of visual preservation in the SC.

Conclusions: : Based on optokinetic testing and SC mapping studies, hESC-RPE cultured on parylene substrates contributed to the preservation of the visual responses in RCS rats.This study suggests that hESC-RPE seeded parylene implants may be considered for treatment strategies to slow the progression of dry-type age-related macular degeneration.

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