Cultured Human Corneal Endothelial Cell Transplantation with a Collagen Sheet as a Substitute Carrier of Descemet’s Membrane

Abstract

Abstract: : Purpose: To evaluate the function of cultured human corneal endothelial cells (HCEC) in vivo and the feasibility of HCEC transplantation with a collagen sheet as the substitute carrier of Descemet’s membrane. Methods: Adult human donor cornea derived from cultured HCEC was labeled with CM–DiI and seeded on a collagen sheet. The pump function of the HCEC sheet was evaluated by measurement of the potential difference and short circuit current. A 6–mm sclerocorneal incision and Descemeto rhexis were performed on rabbit eyes. The HCEC sheet was brought into the anterior chamber and fixed to the posterior stroma (HCEC group). Rabbit corneas with Descemeto rhexis without HCEC sheet transplantation were the controls (control group). Each group, observed for 28 days after surgery, underwent histological and fluorescein microscopic examinations. Results: Pump function parameters of the HCEC sheets were 76–95 % those of human donor corneas. Mean corneal thickness in the HCEC group was significantly less than in the control group 1, 3, 7, 14, 21, and 28 days (p<0.05) after surgery. DiI–labeled cells were spread over the rear corneal surface in the HCEC but not the control group. Marked stromal edema was present in the control group in HE staining, but none in the HCEC group with collagen sheets–bearing monolayer cells. Conclusions: Cultured HCEC from adult human donor cornea have the essential CEC function of maintaining dehydration of the corneal stroma in vivo. Our findings are evidence of the feasibility of transplantation for CEC dysfunction using cultured HCEC with a collagen sheet.

Keywords: cornea: endothelium • transplantation • anterior chamber