| Objective To observe the change of cell proliferative activity and the change of cell cycle after cat corneal endothelial cells (CECs) co-cultured with choroidal melanoma cells (OCM-1). To observe the change of morphology and the changes of centrosomes and cytoskeleton expression of the CECs after co-cultured.Methods The Transwell co-culture systems were divided into three groups: experimental group, positive control group and control group. CECs were co-cultured with OCM-1 cells in the experimental group, CECs were co-cultured with rabbit corneal stromal cells in the positive group, and CECs were cultured alone in the control group. The changes of cat corneal endothelial cells' proliferative activity and 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. The expression of centrosomes were observed byγ-tubulin antibody Centrosome Marker indirect immunofluorescence staining. The expression of cytoskeleton was observed byα-tubulin antibody Cytoskeleton Marker indirect immunofluorescence staining.Results Compared with the control group, the portions of CECs in S-phase of OCM-1 group and stromal-cells group were increased, and the portion of cat endothelial cells in G1-phase of the two groups were decreased. CECs' cell cycle changes in OCM-1 group are more obviously than in stromal-cells group. The portion of CECs in S-phase of OCM-1 group increased nearly 19%, indicating a significant enhance of the cell proliferation. CECs of OCM -1 group growth well, their morphology resembled nature CECs. Centrosomes and cytoskeleton were demonstrated by immunofluorescence staining, their morphology and quantity were normal.Conclusion (1) CECs co-cultured with OCM-1 cells can promote theirs proliferation in certain extent. (2)After CECs co-cultured with OCM-1, their morphology and expression of centrosomes were not significantly different with normal CECs.
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