June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Survival and functionality of iPSC-RPE cultured as a polarized monolayer on ultrathin parylene assessed in a new immunodeficient RCS rat model
Author Affiliations & Notes
  • Biju Thomas
    Ophthalmology, USC Eye Institute, Chino Hills, California, United States
    USC Institute for Biomedical Therapeutics, Los Angeles, California, United States
  • Yousuf Shad
    McMaster University, Hamilton, Ontario, Canada
  • Juan Carlos Martinez
    Ophthalmology, USC Eye Institute, Chino Hills, California, United States
    USC Institute for Biomedical Therapeutics, Los Angeles, California, United States
  • Niharika Singh
    Ophthalmology, USC Eye Institute, Chino Hills, California, United States
  • Young Chang Kim
    Ophthalmology, USC Eye Institute, Chino Hills, California, United States
  • Seth Freeman
    Ophthalmology, USC Eye Institute, Chino Hills, California, United States
  • Sean Anthony Mu
    Ophthalmology, USC Eye Institute, Chino Hills, California, United States
  • Vishweshwer Shastri
    Ophthalmology, USC Eye Institute, Chino Hills, California, United States
  • Kapil Bharti
    National Eye Institute, Bethesda, Maryland, United States
  • Mark S Humayun
    Ophthalmology, USC Eye Institute, Chino Hills, California, United States
    USC Institute for Biomedical Therapeutics, Los Angeles, California, United States
  • David R Hinton
    Pathology, Keck School of Medicine, University of Southern California, Los Angeles, California, United States
  • Danhong Zhu
    Pathology, Keck School of Medicine, University of Southern California, Los Angeles, California, United States
  • Footnotes
    Commercial Relationships   Biju Thomas, None; Yousuf Shad, None; Juan Martinez, None; Niharika Singh, None; Young Chang Kim, None; Seth Freeman, None; Sean Mu, None; Vishweshwer Shastri, None; Kapil Bharti, None; Mark Humayun, None; David Hinton, None; Danhong Zhu, None
  • Footnotes
    Support  Research to Prevent Blindness (RPB), Bright Focus Foundation
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 3263. doi:
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      Biju Thomas, Yousuf Shad, Juan Carlos Martinez, Niharika Singh, Young Chang Kim, Seth Freeman, Sean Anthony Mu, Vishweshwer Shastri, Kapil Bharti, Mark S Humayun, David R Hinton, Danhong Zhu; Survival and functionality of iPSC-RPE cultured as a polarized monolayer on ultrathin parylene assessed in a new immunodeficient RCS rat model. Invest. Ophthalmol. Vis. Sci. 2017;58(8):3263.

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

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Abstract

Purpose : RPE replacement therapy is now evolved as a feasible approach to treat human retinal degeneration diseases like Age-related Macular Degeneration (AMD). This study is aimed to evaluate the survival and functionality of induced pluripotent stem cell derived retinal pigment epithelium (iPSC-RPE) cultured as a polarized monolayer on parylene substrate and transplanted into the subretinal space of a new immunodeficient Royal College of Surgeon (RCS) rat disease model.

Methods : Polarized iPSC-RPE (passage two cells generated from healthy adult human fibroblast cells) was cultured for 2 weeks on rectangular pieces of ultrathin parylene (0.4mm x 0.9mm). After the cells developed into a confluent monolayer, the implants were surgically placed into the subretinal (SR) space of postnatal day (P) 28 RCS rat pups (n=10). Histological assessments were performed using H&E and immunostaining techniques. Visual function was tested using an optokinetic (OKN) head-tracking apparatus and by electrophysiological mapping of the superior colliculus (SC).

Results : iPSC-RPE cultured on ultrathin parylene substrate appeared as a confluent monolayer and expressed RPE 65 and ZO1. In vitro characterization data demonstrated that this iPSC-RPE shows good similarity to fetal RPE (compared to hESC-RPE). Based on histological evaluation at 1 month and 2 month post-implantation, the iPSC-RPE survived and maintained its monolayer structure in all implanted rats. The transplanted RPE expressed RPE 65, iRBP and performed phagocytosis of shed photoreceptor outer segments. Visual functional improvement in immunodeficient dystrophic RCS rats following iPSC-RPE transplantation was demonstrated by SC luminance threshold mapping data (iPSC-RPE implanted rats -3.1±1.7 log cd/m2 vs non-transplanted rats -0.6±0.3 log cd/m2). Based on OKN visual behavioral test, good preservation of visual activity was observed in the iPSC-RPE implanted eyes whereas no such responses were observed in the non-implanted eyes.

Conclusions : iPSC-RPE cells implanted as a polarized monolayer into the SR space of immunodeficient RCS rats can survive as a monolayer and contribute to visual functional benefits. The results support the use of such implants for therapeutic approaches aimed at slowing the progression of outer retinal degeneration diseases.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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