OBJECTIVETo evaluate the feasibility of using cultured corneal endothelial cell (CECs) transplantation for cornea endothelium cell destruction with Gelatin membrane as the carrier in rabbits.

METHODSThe cultured CECs were labeled by Brdu and subcultured in vitro on glutaraldehyde-fixed Gelatin membranes and then the membranes were glued by alpha-cyanoacrylate alkyl to 7.00 mm autologous rabbit corneal bottons whose endothelium were mechanically removed previously. The buttons were sutured in place. With this method the right eyes of 21 rabbits were transplanted with CECs, and the right eyes of another 17 rabbits were transplanted with non-cells carrier as controls. The rabbits were bred and observed by slit microscopy and confocal microscopy at 1, 2, 4, 8, and 12 weeks after surgeries. Also, introcular pressure and corneal thickness were measured by Perkin's tonometer and ultrasonic pachymeter. After 12 weeks, all the animals were sacrificed and the grafts were examined by light microscopy and electronic microscopy.

RESULTSCECs grew well on the gelatine memberane, and formed confluent monolayers in 3-5 days; the cell density reached as high as 2700 cells/mm2. After 2 weeks of operation, all corneal buttons were edema and began to be opaque. The control eyes remained opaque throughout the observation period. In eyes with CECs transplanted, the grafts began to be clear and thin 4 weeks after operation. The cell density of grafts decreased along with time, and the mean cell density of CECs transplantation buttons was (2023.3 +/- 330.3) cells/mm2 12 weeks after operation. The transplanted cells were stained with the anti-Brdu monoclonal antibody.

CONCLUSIONIt is feasible to culture and translate CECs with the Gelatin membrane.