Morphology and Adhesion Complex of Cultured Epithelium, on Amniotic Membrane in Vitro and in Vivo

ABSTRACT

PURPOSE: The morphologic characteristics and adhesion complex formation of cultured limbal epithelium of rabbit on amniotic membrane, in vitro and in vivo. METHODS: Rabbit limbal explants were cultured in vitro on amniotic membrane for 4 weeks. In the in vivo culture, the rabbit corneal epithelium was removed. Next, a tunnel was created at the limbus and, the edge of amniotic membrane was secured in the tunnel and cultured for 4 weeks. The proliferation of epithelium on the amniotic membrane was observed for 4 weeks at 1 week intervals. RESULTS: AE-5 immunohistochemical staining was positive and PAS staining was negative for cultured rabbit limbal epithelium, in vitro and in vivo. Hematoxylin and Eosin staining showed the morphologic characteristics of normal rabbit corneal epithelium in both groups. Transmission electron microscopy performed at an interval of 1 week showed adhesion complex by 3 weeks of in vitro culture, and no significant change was seen until week 4. The formation of the adhesion complex was shown starting at week 1 of in vivo culture and increased until week 4. CONCLUSIONS: The morphology of corneal limbal epithelium of rabbits cultured on amniotic membrane in vitro and in vivo, did not differ significantly compared with normal rabbit epithelium. In vivo culture resulted in more a normal-looking adhesion complex compared with the in vitro culture.