Abstract
Purpose :
Cell therapy is a potential future therapeutic modality for treating corneal endothelial decompensation. We are developing a “ready-to-use” cryopreserved cellular product of human corneal endothelial cells (HCECs). This current study aims to investigate the feasibility of the formation of cryopreserved HCECs using a rabbit model.
Methods :
HCECs were cultured from donor corneas and passaged for 4 times. Cells were then harvested, suspended in a frozen medium supplemented with 100 μM Y-27632, and frozen at -80°C. Eight mm-diameter of the corneal endothelium was denuded in 6 eyes of 6 rabbits. Then, HCECs were thawed at room temperature and injected into the anterior chamber (8.0×105 cells/300μl) followed by 3 hour face-down position. The anterior segment was evaluated by a slit-lamp microscope for 5 days. Immunofluorescence staining of CD166 (human corneal endothelial marker), ZO-1 (barrier function marker), and Na+/K+-ATPase (pump function marker) was performed.
Results :
Slit-lamp microscopy showed the regeneration of corneal transparency in all 6 eyes 5 days after HCECs injection. No local complications were observed, including rejection, severe inflammation, infection, or corneal angiogenesis. Immunofluorescence of flat-mounted corneas showed that CD166-positive hexagonal monolayer corneal endothelium was regenerated in a denuded 8 mm-diameter area, while CD166 was negative in peripheral remaining rabbit corneal endothelium. Regenerated corneal endothelium expressed ZO-1 and Na+/K+-ATPase similar to the normal cornea.
Conclusions :
Current data support the safety and efficacy of our formation of cryopreserved HCECs.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.