Effect Of Rabbit Bone Mesenchymal Stem Cells On Corneal Endothelial Cells Proliferation And Expression Of P21 And PCNA

Objective To observe the proliferative capacity and the changes of cell cycle on rabbit corneal endothelial cells (CECs) which were co-cultured with bone marrow mesenchymal stem cells (BMSCs) in vitro. To explore the mechanism of proliferation on corneal endothelial cells which co-cultured with bone marrow mesenchymal stem cells and assayed the expression of p21 and PCNA.Methods Rabbit corneal endothelial cells and bone marrow mesenchymal stem cells were co-cultured in the Transwell system, the changes of 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 light microscopy. We used Real-time fluorescent quantitative PCR to assay the expression of p21 and PCNA gene of CECs, compared with the experimental group and the control group, statistically analyse the mechanism. of proliferation.Results Compared with the control group, the experimental group showed the portions of CECs in S-phase increased and G1-phase decreased. The portion of CECs in S-phase of experimental group increased 5%, 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. By real-time fluorescent quantitative PCR assay, there is a significant difference of the expression of p21 gene and PCNA gene between the experimental group and the control group on corneal endothelial cells.Conclusion (1) The proliferative capacity of CECs was enhanced in certain extent after co-cultured with bone marrow mesenchymal stem cells in vitro.(2) The cell morphology and the biological behavior of CECs was not significantly different compare with the normal CECs. (3) The probable mechanism of proliferation on corneal endothelial cells after co-cultured with BMSCs was related to downregulation of the expression of p21 gene, and/or upregulation of the expression of PCNA gene.