Cell Sheet Engineering for Primary Corneal Endothelial Cells and Its Animal Model

Human corneal endothelial cells (HCECs) have no proliferative capacity in v^-vo, these cells decrease in number with aging, disease, or trauma. When the cell density drops below a critical level, the corneal endothelial cells can no longer pump enough water to compensate for diffusion into the cornea, resulting in stromal edema, corneal clouding and eventual vision loss. Conventional penetrating keratoplasty involves replacing the entire corneal layer to treat patients whose cornea is healthy except for damaged endothelium. However, access to donated tissue is limited worldwide resulting in critical need for new sources of corneal grafts. The number of such grafts produced by each donor eye could be increased significantly if corneal endothelial cells were expanded in culture before grafting. In this study, we developed a novel therapy technique to fabricate and transplant cultured rabbit corneal endothelial cell (RCECs) sheets for corneal endothelial reconstruction. On the basis of plasma surface modification, we have designed a two-steps method to prepare a y-polyglutamic acid-grafted polyvinylidene fluoride (PVDF) membrane for RCECs cultivation. Confluent cell cultures were detached as a laminated sheet by reductant additive. In vitro characteristics of HCECs sheets were evaluated by histology, immunohistochemistry, and viability. We hope this work may be an optimized method to fabricate bioengineered corneal endothelium and show potential to facilitate corneal endothelial cells transplantation in the future.