Abstract
Purpose :
The aim of this study is to demonstrate the therapeutic outcome of two distinct preparations of functional primary corneal endothelial cells (CECs) for cell injection in a non-human primate (NHP) of corneal endothelial dysfunction.
Methods :
Donor corneas unsuitable for transplantation were procured and processed either by (1) isolation and propagation to the second passage using a dual media cell expansion protocol as Group A cultured CECs, or by (2) utilizing a simple non-cultured endothelial cells (SNEC) harvesting methodology as Group B non-cultured CECs. Two species of NHP (rhesus and cynomolgus) were used in this study.
On the day of transplantation, the corneal endothelium of the macaque was first carefully scrapped. For Group A macaques, at least 6 x 105 of expanded CECs were injected into the anterior chamber. For Group B macaques, at least 1.5 X 105 of SNEC-harvested CECs were used in each injections. Rho-associated protein kinase inhibitor (ROCKi; AR-13503 - 5μM (ARVO-2023)) was supplemented in all procedures. Negative control animals received non-cellular sham injections. All animals were left in a proned, face-down position for a minimum of 3 hours.
Central corneal thickness (CCT) and corneal clarity were assessed weekly for at least 30 days, with some animals maintained for up to 4 months. End-point analysis included immunohistochemistry of human-specific nuclei (HuNu and STEM101) in the excised corneas of the macaques.
Results :
Our results showed that both Group A macaques (receiving expanded CECs) and Group B macaques (receiving SNEC-harvested CECs ), had comparable CCT to that of the pre-operated cornea throughout the main time-points assessed. Corneas in both groups of animals remained clear for at least 30 days, and up to 4 months in animals assigned for long-term evaluation. Corneal endothelium of the excised macaque's corneas receiving cell injections were found to express human specific nuclei, suggesting that functionality was conferred by injected primary CECs in both Group A and Group B macaques. For negative control animals, CCT increased to over 1,000μm by the fourth day, and corneas were opaque throughout the assessment period.
Conclusions :
This current study using NHP showed that both preparations of cadaveric donor-derived CECs are robust in yielding functional primary CECs for corneal endothelial cell injection therapy.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.