June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Histological correlation of superior collicus response to light preserved by transplantation of hESC derived RPE monolayer in RCS rats
Author Affiliations & Notes
  • Biju Thomas
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Padmaja Thomas
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Laura Liu
    Cell and Neurobiology, University of Southern California, Los Angeles, CA
    Ophthalmology, Chang Gung Memorial Hospital, Taoyuan, Taiwan
  • Yuntao Hu
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Danhong Zhu
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
    Pathology, University of Southern California, Los Angeles, CA
  • Dennis Clegg
    Center for Stem Cell Biology and Engineering, University of California-Santa Barbara, Santa Barbara, CA
  • David Hinton
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
    Pathology, University of Southern California, Los Angeles, CA
  • Mark Humayun
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Footnotes
    Commercial Relationships Biju Thomas, None; Padmaja Thomas, None; Laura Liu, None; Yuntao Hu, None; Danhong Zhu, None; Dennis Clegg, Regenerative Patch Technologies (I); David Hinton, RPT (I), RPT (P); Mark Humayun, Bausch & Lomb (F), Bausch & Lomb (C), Bausch & Lomb (P), Bausch & Lomb (R), Bausch & Lomb (S), Alcon (C), Alcon (R), Iridex (P), Iridex (R), Replenish (I), Replenish (C), Replenish (R), Replenish (S), Second Sight (F), Second Sight (I), Second Sight (C), Second Sight (P), Second Sight (R), Second Sight (S), Regenerative Patch Technologies (I), Regenerative Patch Technologies (C)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 1409. doi:
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      Biju Thomas, Padmaja Thomas, Laura Liu, Yuntao Hu, Danhong Zhu, Dennis Clegg, David Hinton, Mark Humayun; Histological correlation of superior collicus response to light preserved by transplantation of hESC derived RPE monolayer in RCS rats. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1409.

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

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Abstract

Purpose: Histological evaluation of the retina of Royal College of Surgeon (RCS) rats that received subretinal implantation of human embryonic stem cell (hESC) derived retinal pigment epithelial (RPE) cells cultured on the biocompatible polymer parylene as a monolayer and its correlation with visual functional preservation in the superior colliculus (SC).

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 postnatal (P) 28 RCS rats (n=16). All animals were maintained under immunosuppression for the entire period of study. Histological evaluation was performed based on immunostaining. In selected animals, electrophysiological mapping of the SC using various light intensity stimulus was performed at 2 months post-implantation.

Results: At P90, immunostaining demonstrated robust photoreceptor (rod and or cone) survival in an area overlying the hESC-RPE implant. In non-treated dystrophic RCS eyes, photoreceptor survival at this age was negligible. Long-term (6 months post-implantation) evaluation confirmed the preservation of rod and cone photoreceptors in the implant area. In all hESC-RPE implanted animals, at threshold stimulus level, responses in the SC could be recorded only from an area corresponding to the implant location in the eye. Immunostaining revealed better rod preservation (4.25±0.59 layers) among rats that responded to low level light stimulation (-5.4 log cd/m2). Rats that responded only at higher light stimulation (>-4.4 log cd/m2) showed less rod survival (2.25±1.1 layers) and majority of these rods were inferior in appearance. No such differences between two luminance measurement groups were observed in the degree of cone survival.

Conclusions: hESC-RPE cultured on parylene substrates implanted into the RCS retina is capable of preserving rod and cone photoreceptors up to P180. Histological correlation of SC luminance measurement suggests preservation of functional rod and cone photoreceptors. The results support the use of such implants for therapeutic approaches aimed at slowing the progression of outer retinal dystrophies such as age-related macular degeneration (AMD).

Keywords: 701 retinal pigment epithelium • 508 electrophysiology: non-clinical • 687 regeneration  
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