March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Morphological and Functional Evaluation of hESC-RPE Cell Suspension Injection in RCS Rats
Author Affiliations & Notes
  • Laura Liu
    Cell and Neurobiology,
    University of Southern California, Los Angeles, California
    Ophthalmology, Chang Gung Memorial Hospital, Taoyuan, Taiwan
  • Yuntao Hu
    Ophthalmology, Doheny Eye Institute, Los Angeles, California
    Ophthalmology, Peking University Third Hospital, Beijing, China
  • Alejandra Gonzalez-Calle
    Ophthalmology, Doheny Eye Institute, Los Angeles, California
  • Danhong Zhu
    Pathology,
    University of Southern California, Los Angeles, California
    Ophthalmology, Doheny Eye Institute, Los Angeles, California
  • Padmaja B. Thomas
    Ophthalmology, Doheny Eye Institute, Los Angeles, California
  • Biju B. Thomas
    Ophthalmology, Doheny Eye Institute, Los Angeles, California
  • Gerald J. Chader
    Ophthalmology, Doheny Eye Institute, Los Angeles, California
  • Dennis O. Clegg
    Center for Stem Cell Biology and Engineering, University of California-Santa Barbara, Santa Barbara, California
  • David R. Hinton
    Pathology,
    University of Southern California, Los Angeles, California
    Ophthalmology, Doheny Eye Institute, Los Angeles, California
  • Mark S. Humayun
    Ophthalmology, Doheny Eye Institute, Los Angeles, California
  • Footnotes
    Commercial Relationships  Laura Liu, None; Yuntao Hu, None; Alejandra Gonzalez-Calle, None; Danhong Zhu, None; Padmaja B. Thomas, None; Biju B. Thomas, None; Gerald J. Chader, 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, 311. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Laura Liu, Yuntao Hu, Alejandra Gonzalez-Calle, Danhong Zhu, Padmaja B. Thomas, Biju B. Thomas, Gerald J. Chader, Dennis O. Clegg, David R. Hinton, Mark S. Humayun; Morphological and Functional Evaluation of hESC-RPE Cell Suspension Injection in RCS Rats. Invest. Ophthalmol. Vis. Sci. 2012;53(14):311. doi: https://doi.org/.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : To evaluate whether transplantation of retinal pigment epithelial (RPE) cells derived from H9 human embryonic stem cells (hESC-RPE) delivered as cell suspension can maintain the RPE functionality and prevent photoreceptor degeneration in the dystrophic Royal College of Surgeons (RCS) rat. The study will also evaluate the immunological reactions of the host retina to non-polarized hESC-RPE.

Methods: : HESC-RPE cells were cultured for 4 weeks and digested into cell suspension at the cell density of 5.0x107/ml. Subretinal injections of hESC-RPE cells (105, 2µl) were performed in 28 to 31 day old RCS rats. Prednisolone was administered to all rats through drinking water (0.002mg/liter) for the entire period of study. Animals were sacrificed and eyes enucleated at 10 days (n=3), and 2 months (n=9) after surgery; tissue sections underwent histological evaluation using hemotoxylin and eosin (H&E) staining and immunostaining for RPE markers (RPE65), human marker (TRA-1-85), and immune response markers (CD3, CD68, GFAP). Visual functional evaluation was performed using optokinetic testing. Aperio Scanscope was used for measurement of outer nuclear layer thickness and performing cell counts.

Results: : Based on H&E staining, presence of pigmented cells was observed in all transplanted rats. In the 10 days post-transplantation group, hESC-RPE cells showed co-localized expression of TRA-1-85 and RPE-65 indicative of viable donor cells. In this group, transplanted cell clumps were surrounded by CD68 positive cells indicating macrophage activity. In the 2 months post-transplantation group, CD68 expression was observed mostly over the cell clumps, but TRA-1-85 and RPE-65 markers were not identified. Optokinetic testing suggested some improvement in the implanted eye; however the difference with the non-transplanted eye was not significant. Based on histological assays, no appreciable rescue of the outer retinal layer was observed in the transplanted area.

Conclusions: : Dissociated hESC-RPE cells do not survive long-term and do not function in the rat’s subretinal space. One possible mechanism for the accelerated loss of the transplanted hESC-RPE cells can be due to the increased host immune reaction to non-polarized hESC-RPE.

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

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×