June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Long-term Efficacy and Safety of an RPE Cell Monolayer Derived From Human Embryonic Stem Cells Implanted into the Subretinal Space of the RCS Rat
Author Affiliations & Notes
  • Ramiro Ribeiro
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, FEPAR, Curitiba, Brazil
  • Michael Koss
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, Goethe University, Frankfurt, Germany
  • Bruno Diniz
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, Federal University of São Paulo, São Paulo, Brazil
  • Rodrigo Brantfernandes
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, Federal University of São Paulo, São Paulo, Brazil
  • Laura Liu
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, Chang Gung Memorial Hospital, Taoyuan, Taiwan
  • Padmaja Thomas
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Biju Thomas
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, CA
  • Gerald Chader
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, CA
  • David Hinton
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, CA
  • Mark Humayun
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, CA
  • Footnotes
    Commercial Relationships Ramiro Ribeiro, None; Michael Koss, None; Bruno Diniz, None; Rodrigo Brantfernandes, None; Laura Liu, None; Padmaja Thomas, None; Biju Thomas, None; Gerald Chader, None; 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, 2219. doi:
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      Ramiro Ribeiro, Michael Koss, Bruno Diniz, Rodrigo Brantfernandes, Laura Liu, Padmaja Thomas, Biju Thomas, Gerald Chader, David Hinton, Mark Humayun; Long-term Efficacy and Safety of an RPE Cell Monolayer Derived From Human Embryonic Stem Cells Implanted into the Subretinal Space of the RCS Rat. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2219.

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

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Abstract
 
Purpose
 

To evaluate long term safety and efficacy of RPE derived from human embryonic stem cells (hESCs) seeded as a monolayer on a sub-micron parylene-C membrane implanted into the subretinal space of the dystrophic Royal College of Surgeons (RCS) rat

 
Methods
 

All experiments were performed in compliance with the ARVO Statement on the Use of Animals Care and Use Committee (IACUC) at the University of Southern California. A mesh-supported submicron parylene-C membrane was seeded with hESC-RPE, differentiated into a polarized monolayer, and implanted in the RCS rats (n=20). A sham group (n=10) received the implant of the parylene membrane without cells. The fellow eye (n=30) served as the untreated control group. Visual behavior was measured at different time points using optokinetic (OKN) head tracking. Eyes were processed for histology at P150, P210, P270 and P390. An experienced pathologist analyzed the retinal morphology. The outer nuclear layer (ONL) was quantified using an Aperio scan scope. Characterization of human RPE was performed using a human-specific cell surface marker (anti-TRA-185) and anti-RPE65. Rhodopsin and cone markers were used to demonstrate photoreceptor rescue

 
Results
 

Animals implanted with hESC-RPE performed significantly better (p<0.05) than the sham and control groups on the OKN behavioral test after P63. For example, at P77 the head tracking results in the hESC-RPE implanted group averaged 16.07 ± 0.33 seconds, the sham group 11 ± 0.84 seconds and the non-operated group 8.30 ± 0.16 seconds. Histologic studies did not show cell migration off the membrane or tumor formation. Nuclei in the ONLs were in higher number in the implanted (183 ± 16 cells) than the non-treated (15 ± 6 cells) eye at P150. RPE65, TRA-185, rhodopsin and cone markers were positive in the implant site of the majority of the rats implanted with hESC-RPE but not in control/sham cases receiving no hESC-RPE cells

 
Conclusions
 

The transplantation of an hESC-RPE monolayer seeded on a membrane was safely performed and was associated with a better long term functional outcome than with the sham and control groups

 
 
RCS rat retina implanted with hESC-RPE seeded on a parylene membrane
 
RCS rat retina implanted with hESC-RPE seeded on a parylene membrane
 
Keywords: 721 stem cells • 695 retinal degenerations: cell biology • 412 age-related macular degeneration  
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