Comparison Of The Proliferative Capacity Of Cat Corneal Endothelial From The Central And Peripheral Areas

Purpose To compare the proliferative capacity between corneal endothelial cells from the peripheral and central areas of cat corneal.To observe the change of cell proliferative capability and the change of cell cycle after cat corneal endothelial cells(CECs) co-cultured with glioma cells(U251),then make the preliminary evaluation of biological property of co-cultured CECs.Methods Cat corneas were divided into younger group and older group based on cat's age.We use Real- time fluorescent quantitative PCR to assay the telomerase activity of CECs which were divided into central(0-6.0mm) and peripheral areas(6.0-9.5mm) by the use of trephines.The potential proliferative capacity of CECs both in peripheral and central areas of cornea was investigated.CECs were divided into three groups for the purpose of investigating the factors which promote the CECs' proliferation.CECs were co-cultured with U251 cells in the experimental group and with cat corneal stromal cells in the positive group by transwell systems, and CECs were cultured alone in the control group.The changes of cat corneal endothelial cell cycle were examined by flow cytometry after co-cultured.The morphological changes of cells before and after co-cultured were observed by inverted phase contrast microscope and HE staining,meanwhile,the telomerase activity of CECs were detected by real-time fluorescent quantitative PCR after co-cultured.Results There is a significant difference of the telomerase activity between the central and peripheral areas(F=58.51,P=0.000) and no statistically significant difference between the younger and older age group(F=6.21,P=0.028) in cats'corneal endothelium.Compared with the control group,both the experimental group and positive group showed the portions of CECs in S-phase increased and G1-phase decreased,which is more obvious in experimental group.The portion of CECs in S-phase of experimental group increased nearly 13%,indicating a significant enhancement of the cell proliferative capacity.After co-cultured,the number of CECs in experimental group has increased,there's no obviously abnormality of CECs morphologically compared with normal CECs.Telomerase activity of CECs in the experimental group increased in different degree compared with those in the positive group and the control group,but it is far less than those of the glioma cells.Conclusion(1) cat corneal endothelial cells from peripheral areas may be exist stem-like cells and retain a higher potential proliferative capacity which does not related with the age.(2). After co-cultured with U251 cells,the proliferative capacity of CECs shows a significant enhancement,the biological behavior of CECs was not significantly different compare with the normal CECs.