[Construction of corneal endothelium deficiency model by cryo-injury and its application].

Objective: To set up an animal model of corneal endothelium deficiency and test the bio-safety and pump function of B4G12 cells with this model.

Methods: Thirty SD rats were divided into cryo-injury group and cryo-injury with B4G12 cell transplantation group. Models of corneal endothelium deficiency were created by cryo-injury with liquid nitrogen, and then B4G12 cells were transplanted into the eyes by anterior chamber injection. Corneas were checked under slit lamp and confocal microscope observations at some specific time points, pathological staining was also performed.

Results: The corneal endothelial cells were removed completely from the Descemet's membrane after cryo-injury. Cells transplanted firmly stuck to the Descemet's membrane. The corneas in the cryo-injury group were swelling with haze, while in the transplantation groups, the corneas restored transparence and normal thickness 2 weeks after the surgery. Confocal microscope and HE staining confirmed that in the cryo-injury group, the Descemet's membrane was denuded and the cornea stroma layer was in edema, on the contrary, in the other group, the transplanted cells completely covered the Descemet's membrane.

Conclusion: Cryo-injury can be used for building the rat model of corneal endothelium deficiency, no neoplasm and inflammation reaction were found during the observation after B4G12 cells transplanted and B4G12 cells had the pump function in vivo.