AIM:To evaluate chondroitin sulfate on the glycerol cryopreservation of corneal endothelial cells(CEC).METHODS:Forty Wistar rats corneal pieces were divided into the experimental group:glycerol cryopreservation medium containing chondroitin sulfate(CS);and the control group:mere glycerol cryopre-servation medium.The two groups were saved at-20℃ for 2 months.All pieces were examined with trypan blue combined with alizarin red staining(viability staining) and electron-microscope examination for CEC.30 SD rats were taken to establish Wistar-SD rat model of allogenic penetrating keratoplasty.Slit lamp microscope detected the occurrence of corneal rejection,TNF-α and IFN-γ expression was examined by immunohistochemistry to test the effect of preservation.RESULTS:The staining results,electron-microscope examination and TNF-α,IFN-γ expression showed,the difference was significant in these two groups(P<0.05),the results of the experimental group was better than the control group after the saving and the corneal transplantation(P<0.05).The electron-microscope examination showed indivdual vacuoles in CEC cytoplasm,slight swelling of mitochondrial and endoplasmic reticulum in experimental group,obvious swelling of CEC mitochondrial and endoplasmic reticulum in control group.CONCLUSION:Glycerol cryopreservation can preserve the viability of CEC.Chondroitin sulfate can obviously improve the effect of preservation.