June 2020
Volume 61, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2020
hESC-derived retina organoids produced by a scalable cGMP compatible process improve visual function after transplantation to immunodeficient RD rats
Author Affiliations & Notes
  • Bin Lin
    Stem Cell Research Center, University of California at Irvine, Irvine, California, United States
  • Bryce McLelland
    AIVITA Biomedical, Irvine, California, United States
  • Yuntian Xue
    Biomedical Engineering, University of California at Irvine, Irvine, California, United States
    Stem Cell Research Center, University of California at Irvine, Irvine, California, United States
  • Robert Sims
    Stem Cell Research Center, University of California at Irvine, Irvine, California, United States
  • Johanes Santoso
    Stem Cell Research Center, University of California at Irvine, Irvine, California, United States
  • Gabriel Nistor
    AIVITA Biomedical, Irvine, California, United States
  • Robert Aramant
    Stem Cell Research Center, University of California at Irvine, Irvine, California, United States
  • Andrew Browne
    Ophthalmology, Gavin Herbert Eye Institute, University of California at Irvine, Irvine, California, United States
    Institute for Clinical and Translational Science, University of California at Irvine, Irvine, California, United States
  • Hans Keirstead
    AIVITA Biomedical, Irvine, California, United States
  • Magdalene J Seiler
    PM&R, Ophthalmology, Anatomy & Neurobiology, University of California at Irvine, Irvine, California, United States
    Stem Cell Research Center, University of California at Irvine, Irvine, California, United States
  • Footnotes
    Commercial Relationships   Bin Lin, None; Bryce McLelland, AIVITA Biomedical (E); Yuntian Xue, None; Robert Sims, 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, RPB unrestricted grant to UCI Department of Ophthalmology, ICTS KL2 Grant number is KL2 TR001416
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 2505. doi:
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      Bin Lin, Bryce McLelland, Yuntian Xue, Robert Sims, Johanes Santoso, Gabriel Nistor, Robert Aramant, Andrew Browne, Hans Keirstead, Magdalene J Seiler; hESC-derived retina organoids produced by a scalable cGMP compatible process improve visual function after transplantation to immunodeficient RD rats. Invest. Ophthalmol. Vis. Sci. 2020;61(7):2505.

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

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Abstract

Purpose : The development of human embryonic stem cell (hESC) derived retina organoids produced by a scalable cGMP compatible process was followed after transplantation to the subretinal space of immunodeficient and immunocompetent, immunosuppressed rho S334ter-3 (RD) rats.

Methods : A scalable cGMP compatible process was established for the generation and characterization of a Working Cell Bank (WCB) of CSC-14 hESCs (NIH 0284). hESCs were differentiated into retina organoids and characterized by immunohistochemistry (IHC), flow cytometry and qPCR. Two-Photon microscopy was used for functional and structural imaging of organoids by fluorescence lifetime imaging microscopy (FLIM) and hyperspectral imaging (HSpec).
Retina organoid sheets were transplanted to the subretinal space of RD hosts (P31-51). Transplants were monitored by Optical Coherence Tomography (OCT). In immunocompetent RD rats, immunosuppressant (tacrolimus and mycophenolate mofetil) levels in blood were determined by LC-MS. Cytostatic effects on target lymphocyte populations was evaluated by Flow Cytometry.
Visual function was accessed by optokinetic tests (OKT) and superior colliculus (SC) electrophysiology. Sections through transplants were stained with hematoxylin & eosin (H&E), or processed for IHC to label human donor cells, retinal cell types and synaptic markers.

Results : The WCB of CSC-14 hESCs was characterized on viability; Identity (Oct4); karyotype; sterility and neural differentiation potential, showing the WCB meets the FDA requirements.
IHC of retina organoids shows early lamination and development of retinal cell progenitors. Selected organoids for transplantation showed lamination. OCT revealed transplant development and photoreceptor rosettes. Transplanted eyes showed vision improvement by OKT and SC recording. Transplant developed different retinal cells including photoreceptors; and integrated with the host retina. In immunosuppressed rats, therapeutic levels of immunosuppressant remained in the blood and transplants survived after transplantation.

Conclusions : A WCB of CSC-14 hESCs was successfully established and met FDA requirements, using a scalable cGMP compatible process. Retinal organoids mature further after transplantation, develop photoreceptors, integrate into the host retina, and improve visual function.

This is a 2020 ARVO Annual Meeting abstract.

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