Simulation of blinking for the reconstruction of rabbit corneal epithelium on a lyophilized amniotic membrane

In this study, rabbit corneal epithelium was reconstructed using blinking dynamic culture with a selfmanufactured amniotic membrane (AM) supporter composed of three Teflon rings and a lyophilized amniotic membrane (LAM). The reconstructed corneal epithelium was characterized by histological (H and E) and immunohistochemical staining (PCNA) and by RT-PCR and TEM. The basal layer of the reconstructed corneal epithelium was well formed, and the epithelium was tightly constructed due to the medium supply during the dynamic culture. The cytokeratin 3 (CK3) mRNA expression levels in tilting and blinking dynamic cultured third passage RCE cells seeded onto a LAM were about 2 times greater than that in static culture. The morphology of the corneal epithelium reconstructed on a LAM via blinking dynamic culture over a period of six days better resembled that of the normal cornea than the corneal epithelium reconstructed via the static and tilting dynamic culture. Thus, the reconstruction of the corneal epithelium using a LAM and blinking dynamic culture method is considered to be a good in vitro model for the autologous or allogeneic transplantation of corneal epithelium and it can be quite useful in toxicological test kits.