June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
In Vivo Functional Comparison of Polarized and Non-polarized hESC-RPE Cells Transplantation in RCS Rats
Author Affiliations & Notes
  • Laura Liu
    Cell and Neurobiology, University of Southern California, Los Angeles, CA
    Ophthalmology, Chang Gung Memorial Hospitial, Taoyuan, Taiwan
  • Biju Thomas
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Ramiro Ribeiro
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Alejandra Gonzalez-Calle
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Danhong Zhu
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
    Pathology, University of Southern California, Los Angeles, CA
  • Padmaja Thomas
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Keng-Hung Lin
    Ophthalmology, Doheny Eye Institute, Los Angeles, CA
  • Yuntao Hu
    Ophthalmology, Doheny Eye Institute, Los Angeles, 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 Laura Liu, None; Biju Thomas, None; Ramiro Ribeiro, None; Alejandra Gonzalez-Calle, None; Danhong Zhu, None; Padmaja Thomas, None; Keng-Hung Lin, None; Yuntao Hu, 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, 1401. doi:
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      Laura Liu, Biju Thomas, Ramiro Ribeiro, Alejandra Gonzalez-Calle, Danhong Zhu, Padmaja Thomas, Keng-Hung Lin, Yuntao Hu, David Hinton, Mark Humayun; In Vivo Functional Comparison of Polarized and Non-polarized hESC-RPE Cells Transplantation in RCS Rats. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1401.

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

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Abstract

Purpose: To evaluate the visual functional differences in dystrophic Royal College of Surgeons (RCS) rats after transplantation of retinal pigment epithelial cells derived from H9 human embryonic stem cells (hESC-RPE) delivered as a polarized monolayer cell sheet on a thin parylene membrane or non- polarized cell suspension.

Methods: For transplantation of polarized monolayer cell sheet, hESC-RPE cells were cultured for 3 weeks on parylene sheets; for transplantation of cell suspension, hESC-RPE cells were cultured for 3 weeks and digested into cell a suspension before implantation. Subretinal implantation of hESC-RPE /parylene (approximately 3000 cells/implant, n=6) and subretinal injection of hESC-RPE cells (10 5/2µl, n=6) was performed in 27 to 29 day old RCS rats. Six non-implanted RCS rats served as controls. Prednisolone was administered through drinking water (0.002mg/liter) for the entire period of study. Post-surgical evaluation was performed using spectral-domain optical coherence tomography (SD-OCT). Optokinetic head tracking (OHT) testing and luminance threshold measurement from the superior colliculus (SC) were employed for visual functional assessments at 2 months post-surgery.

Results: Based on SD-OCT, preservation of the retinal layers in the transplant area was observed in both implant groups. In hESC-RPE/parylene implanted animals, the head tracking duration was higher (4.06±0.76 sec/min) compared to the suspension injection group (2.56±0.53 sec/min). SC recording demonstrated significantly lower (p=0.008) luminance threshold in the hESC-RPE/parylene implant group (-5.15±0.27 log cd/m2) compared to the subretinal injection group (-3.31±0.49 log cd/m2). SC luminance threshold map showed good correlation with the location of the hESC-RPE/parylene in the retina.

Conclusions: hESC-RPE when implanted as a monolayer or injected as cell suspension demonstrated preservation of the retina in RCS rats up to 2 months post-surgery. Better visual preservation was observed in the hESC-RPE/parylene implanted animals as evidenced by SC luminance threshold measurement and OHT testing. Our study suggests that implantation of hESC-RPE/parylene may be a more desirable therapeutic approach for disease conditions related to RPE dysfunction such as dry age-related macular degeneration.

Keywords: 701 retinal pigment epithelium • 687 regeneration • 412 age-related macular degeneration  
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