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Objective:To explore mechanism of nduction of fibroblast to corneal endothelial cell.Methods:Rabbit conjunctiva fibroblasts were used as feeder cells,rabbit oral mucosa epithelial cells were used as seed cells,and human denuded amniotic membrane was used as carrier to establish tissue engineering corneal endothelium.The transformation effect was observed.Results:As concentration of mitomycin C increased,cell survival rale gradually decreased,cell proliferation was obviously inhibited when concentration ≥25μg/mL;5 days alter being treated by 5μg/mL.mitomycin C,cell body was enlarged and extended without cell fusion,however after being treated by 0.5 μg/mL mitomycin C,cell body was significantly proliferated and gradually fused:alter 3 weeks of culture,stratified epithelium appeared on rabbit oral mucosa epithelial cells,differentiation layers were 4-5 and were well differentiated,the morphology was similar to corneal endothelial cells;Under electron microscope,surface layer of cells were polygonal,tightly connected to another with microvilli on the bonier,there was hemidesmosome between basal cells and human denuded amniotic membrane.Conclusions:Fibroblast cells have the potential of multi-directional differentiation,effective induction can promote emergence of intercellular desmosomes between seed cells and emergence ol epithelial surface microvilli,and differentiate to the corneal endothelial cell.However,clinical application still needs more research and safetv evaluation.
Objective: To explore mechanism of nduction of fibroblast to corneal endothelial cell. Methods: Rabbit conjunctiva fibroblasts were used as feeder cells, rabbit oral mucosa epithelial cells were used as seed cells, and human denuded amniotic membrane was used as carrier to establish tissue engineering corneal endothelium. The transformation effect was observed. Results: As concentration of mitomycin C increased, cell survival rale gradually decreased, cell proliferation was significantly inhibited when concentration ≥ 25 μg / mL; 5 days alter being treated by 5 μg / mL. was enlarged and extended without cell fusion, however after being treated by 0.5 μg / mL mitomycin C, cell body was significantly proliferated and fused: alter 3 weeks of culture, stratified epithelium was found on rabbit oral mucosa epithelial cells, differentiation layers were 4- 5 and were well differentiated, the morphology was similar to corneal endothelial cells; Under electron microscope, surface layer of cell s were polygonal, tightly connected to another with microvilli on the bonier, there was hemidesmosome between basal cells and human denuded amniotic membrane. Conclusions: Fibroblast cells have the potential of multi-directional differentiation, effective induction can promote emergence of intercellular desmosomes between seed cells and emerging ol epithelial surface microvilli, and differentiate to the corneal endothelial cell. However, clinical application still needs more research and safetv evaluation.