June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
Effect of immunosuppression on hESC-derived retina organoids in vitro and in vivo
Author Affiliations & Notes
  • Bin Lin
    Sue and Bill Gross Stem Cell Research Center, University of California Irvine, Irvine, California, United States
  • Robert Sims
    Sue and Bill Gross Stem Cell Research Center, University of California Irvine, Irvine, California, United States
  • Yuntian Xue
    Sue and Bill Gross Stem Cell Research Center, University of California Irvine, Irvine, California, United States
    Biomedical Engineering, University of California Irvine, Irvine, California, United States
  • Bryce McLelland
    AIVITA Biomedical, Irvine, California, United States
  • Raghda Taha Fouda
    Sue and Bill Gross Stem Cell Research Center, University of California Irvine, Irvine, California, United States
  • Johanes Alvian Santoso
    Sue and Bill Gross Stem Cell Research Center, University of California Irvine, Irvine, California, United States
  • Gabriel Nistor
    AIVITA Biomedical, Irvine, California, United States
  • Robert Aramant
    Sue and Bill Gross Stem Cell Research Center, University of California Irvine, Irvine, California, United States
  • Andrew W Browne
    Biomedical Engineering, University of California Irvine, Irvine, California, United States
    Institute for Clinical and Translational Science, and Ophthalmology, Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • Hans Keirstead
    AIVITA Biomedical, Irvine, California, United States
  • Magdalene J Seiler
    Sue and Bill Gross Stem Cell Research Center, University of California Irvine, Irvine, California, United States
    PM&R, Ophthalmology, Anatomy & Neurobiology, University of California Irvine, Irvine, California, United States
  • Footnotes
    Commercial Relationships   Bin Lin, None; Robert Sims, None; Yuntian Xue, None; Bryce McLelland, AIVITA Biomedical (E); Raghda Fouda, None; Johanes Santoso, None; Gabriel Nistor, AIVITA Biomedical (E); Robert Aramant, Ocular Transplantation LLC (E), Ocular Transplantation LLC (P); Andrew Browne, None; Hans Keirstead, AIVITA Biomedical (E), AIVITA Biomedical (S); Magdalene Seiler, Ocular Transplantation LLC (P)
  • Footnotes
    Support  CIRM TR1-10995
Investigative Ophthalmology & Visual Science June 2021, Vol.62, 3297. doi:
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    • Get Citation

      Bin Lin, Robert Sims, Yuntian Xue, Bryce McLelland, Raghda Taha Fouda, Johanes Alvian Santoso, Gabriel Nistor, Robert Aramant, Andrew W Browne, Hans Keirstead, Magdalene J Seiler; Effect of immunosuppression on hESC-derived retina organoids in vitro and in vivo. Invest. Ophthalmol. Vis. Sci. 2021;62(8):3297.

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

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Abstract

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

Methods : A Working Cell Bank (WCB) of CSC-14 hESCs (NIH 0284) was established using a scalable cGMP compatible process. ROs differentiated from hESCs were characterized by immunohistochemistry (IHC), flow cytometry, qPCR and a two-way mixed lymphocyte reaction (MLR). Functional and structural imaging of organoids was obtained using fluorescence lifetime imaging microscopy (FLIM) and hyperspectral imaging (HSpec). Retina organoid sheets were transplanted to the RD hosts and monitored by Optical Coherence Tomography (OCT). Immunosuppressants (20-30mg Tacrolimus [TAC] pellet implant in combination with oral mycophenolate mofetil [MMF]) were applied to the rats. TAC levels in blood were determined by LC-MS. Cytostatic effects on target lymphocyte populations were evaluated by flow cytometry. Visual function was accessed by optokinetic tests and superior colliculus electrophysiology. Sections through transplants were stained with hematoxylin & eosin (H&E) and IHC. The effect of TAC (3 ng/ml)) and MPA (0.5 µg/ml) on retinal organoids was tested by FLIM after 1 and 4 weeks exposure.

Results : The WCB of hESCs meets the FDA requirements. 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. At 2-3 mo. of differentiation, organoids switched their metabolic status from more glycolytic to more oxidative which remained stable at 4 months. Immunosuppressants TAC and MPA showed no influence on the metabolic activity of retinal organoids after 1 and 4 weeks of exposure. OCT revealed transplant development and photoreceptor rosettes. The transplants developed different retinal cells including photoreceptors; and integrated with the host retina. Therapeutic levels of immunosuppressant remained in the blood and helped transplants survive in the host.

Conclusions : Retina organoids matured and developed photoreceptors in long-term culture and after transplantation. Immunosuppressants did not measurably alter the metabolic status of organoids, and prevented rejection of retinal organoid transplants.

This is a 2021 ARVO Annual Meeting abstract.

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