Histological assessment of corneal endothelium regenerated by injection of RCECs and HCECs after preservation in CTV.
(A) Regenerated corneal endothelium was evaluated by immunofluorescent staining 2 weeks after cell transplantation. The function-related markers Na+/K+-ATPase, ZO-1, and N-cadherin were expressed in the corneal endothelium of the eyes injected with RCECs after 24 hours of preservation. Actin similar to that seen in healthy cells was observed along the cell cortex in the eyes injected with RCECs. Fibroblastic cells without function-related markers were observed. Scale bar: 50 μm. (B) Feasibility of using HCECs after preservation in CTV was evaluated by injecting HCECs preserved for 24 hours in CTV into the rabbit corneal endothelial dysfunction model (n = 3). Immunofluorescence staining showed that corneal endothelium was regenerated by the injected HCECs and expressed Na+/K+-ATPase and N-cadherin. Actin staining showed that the regenerated corneal endothelium in the rabbit eyes had a normal morphology consisting of a monolayer of hexagonal cells. Scale bar: 500 μm.