June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
Pluripotent Stem Cell-derived Corneal Endothelial Cells for Donor-Free Keratoplasty
Author Affiliations & Notes
  • Muhammad Ali
    The Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States
  • Shahid Y Khan
    The Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States
  • John Gottsch
    The Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States
  • Eric Hutchinson
    Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States
  • Aisha Khan
    Interdisciplinary Stem Cell Institute, University of Miami, Miami, Florida, United States
  • S. Amer Riazuddin
    The Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States
  • Footnotes
    Commercial Relationships   Muhammad Ali, None; Shahid Khan, None; John Gottsch, None; Eric Hutchinson, None; Aisha Khan, None; S. Amer Riazuddin, None
  • Footnotes
    Support  Maryland Stem Cell Research Fund
Investigative Ophthalmology & Visual Science June 2021, Vol.62, 953. doi:
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      Muhammad Ali, Shahid Y Khan, John Gottsch, Eric Hutchinson, Aisha Khan, S. Amer Riazuddin; Pluripotent Stem Cell-derived Corneal Endothelial Cells for Donor-Free Keratoplasty. Invest. Ophthalmol. Vis. Sci. 2021;62(8):953.

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

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Abstract

Purpose : Corneal endothelial dystrophies are responsible for a large percentage of corneal transplants performed around the world. A number of corneal transplant techniques, full and partial thickness keratoplasties, have been developed that are highly effective in restoring vision. However, in many areas of the world, there is a shortage of transplantable-grade donor corneal tissue. Here, we evaluate the efficacy of cryopreserved human embryonic stem cell (hESC)-derived corneal endothelial cells (CECs) to form a functional monolayer of corneal endothelium (CE) in pre-clinical animal models.

Methods : H9 hESCs were differentiated to generate CECs under xeno-free conditions. A silicone needle was used to remove the resident CECs in an 8 mm diameter area of the central cornea of rabbits and monkeys, and cryopreserved hESC-derived CECs were injected in the anterior chamber. The animals were placed in an eye-down position for three hours to allow the injected cells to settle on denuded Descemets Membrane (DM). The clinical examination of post-operative eyes was performed by evaluating intraocular pressure, corneal thickness, and CEC density at different time points. Immunohistochemical (IHC) analysis was performed to confirm the expression of ZO-1, Na+/K+ ATPase α1, and N-cadherin. A detailed necropsy and CBC/blood chemistry examination was completed 9- and 12-months post-injection in rabbits and monkeys, respectively.

Results : The injected corneas became clear within 3-4 weeks post-injection in both rabbits and monkeys accompanied by a negligible difference in the corneal thickness between the injected (right) eye and the untreated (left) eye. Confocal scanning microscopy confirmed an intact layer of hexagonal/polygonal cells in the CE formed from the cryopreserved hESC-derived CECs. IHC analysis illustrated intact tight junctions, pump function, and structural integrity of the injected cryopreserved hESC-derived CECs. Finally, all parameters examined during the CBC/blood chemistry analysis were within the normal physiological range while necropsy examination confirmed no remarkable change in multiple tissues examined for teratoma formation.

Conclusions : We present an alternative to eye bank donor corneal tissue of cryopreserved hESC-derived CECs for restoring vision in corneal endothelial dystrophies.

This is a 2021 ARVO Annual Meeting abstract.

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