June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Effect of immunosuppression on transplants of hESC-derived retina organoids to retinal degenerate rats
Author Affiliations & Notes
  • Bin Lin
    Stem Cell Research Center, University of California Irvine, Irvine, California, United States
  • Robert Sims
    Stem Cell Research Center, University of California Irvine, Irvine, California, United States
  • Yuntian Xue
    Biomedical Engineering, University of California Irvine, Irvine, California, United States
  • Raghda Taha Fouda
    Stem Cell Research Center, University of California Irvine, Irvine, California, United States
  • Jeffrey Delgado
    Stem Cell Research Center, University of California Irvine, Irvine, California, United States
  • Bryce McLelland
    AIVITA Biomedical, Irvine, California, United States
  • Gabriel Nistor
    AIVITA Biomedical, Irvine, California, United States
  • Hans Keirstead
    AIVITA Biomedical, Irvine, California, United States
  • Andrew Browne
    Biomedical Engineering, Ophthalmology, Gavin Herbert Eye Institute, Institute for Clinical and Translational Science, University of California Irvine, Irvine, California, United States
  • Magdalene J Seiler
    Stem Cell Research Center, University of California Irvine, Irvine, California, United States
    PM&R, Ophthalmology, Anatomy & Neurobiology, University of California at Irvine, Irvine, California, United States
  • Footnotes
    Commercial Relationships   Bin Lin None; Robert Sims None; Yuntian Xue None; Raghda Fouda None; Jeffrey Delgado None; Bryce McLelland AIVITA Biomedical, Code E (Employment); Gabriel Nistor AIVITA Biomedical, Code E (Employment); Hans Keirstead AIVITA Biomedical, Code E (Employment), AIVITA Biomedical, Code S (non-remunerative); Andrew Browne None; Magdalene Seiler Ocular Transplantation LLC, Code P (Patent)
  • Footnotes
    Support  CIRM TR1-10995, RPB unrestricted grant to UCI Department of Ophthalmology, ICTS KL2 Grant number is KL2 TR001416, NIH R01 EY031834
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 4514 – F0301. doi:
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    • Get Citation

      Bin Lin, Robert Sims, Yuntian Xue, Raghda Taha Fouda, Jeffrey Delgado, Bryce McLelland, Gabriel Nistor, Hans Keirstead, Andrew Browne, Magdalene J Seiler; Effect of immunosuppression on transplants of hESC-derived retina organoids to retinal degenerate rats. Invest. Ophthalmol. Vis. Sci. 2022;63(7):4514 – F0301.

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

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Abstract

Purpose : The effect of immunosuppression on the survival of human embryonic stem cell (hESC) derived retina organoids (ROs) produced by a scalable cGMP compatible process was studied after transplantation to the subretinal space of Rho S334ter-3 retinal degeneration (RD) rats, and in vitro.

Methods : A Working Cell Bank (WCB) of CSC-14 hESCs (NIH 0284) was established using a scalable cGMP compatible process. RO sheets differentiated from hESCs were transplanted to 3 groups of rats: 1. Immunodeficient (nude) RD rats; 2. Immunocompetent RD rats; 3. Immunocompetent RD rats with immunosuppression. The survival of the transplants was monitored by Optical Coherence Tomography (OCT). Immunosuppressants (Tacrolimus [TAC] pellet implant and oral mycophenolate mofetil [MMF]) were applied to the rats in group 3. TAC levels in blood were determined by LC-MS and MMF/cytokine levels were monitored by Elisa. Cytostatic effects on target lymphocyte populations were evaluated by flow cytometry. Sections through transplants were stained with hematoxylin & eosin (H&E) and immunohistochemistry (IHC) with different markers. The effect of TAC and MPA on RO was tested by fluorescence lifetime imaging (FLIM) after exposure for 1 and 4 weeks. Visual function was tested by optokinetic testing (OKT).

Results : A WCB of hESCs was established from the MCB. In vitro immunogenicity tests showed that ROs are not likely to induce an immune response. IHC of ROs shows early lamination and development of retinal cell progenitors. No significant difference was found between ROs exposed to immunosuppressants and controls. The transplants developed different retinal cells including photoreceptors; integrated with the host retina; and survived for more than 6 months in immunocompromised rats. CD8 T-cells obliterated donor cells in immune competent rats without immunosuppression after 2 months post-surgery. Therapeutic levels of immunosuppressant remained in the blood and helped transplants survive in the host for 6 months. OKT showed improvement of visual function in transplanted rats compared to non-surgery controls.

Conclusions :
Retina organoids matured, developed photoreceptors, and improved visual acuity after transplantation. Suppression of host CD8 T-cells may be necessary for transplant long term survival. Activity of microglia and macrophages alone may not be detrimental to transplant survival.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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