Morphologic characteristics and formation of adhesion complex of cultured rabbit limbal epithelium on amniotic membrane in vitro

To investigate the morphologic characteristics and the formation of adhesion complex formation of cultured rabbit limbal epithelium on amniotic membrane in vitro.The limbal explants (size: 2 mm x 2 mm) were taken from 15 rabbits and were treated with 1.2 U/ml dispase II at 37 degrees C in 5% CO(2) incubator for 20 min, then, cultured on amniotic membrane in vitro for 4 weeks. It was cultured in culture medium during the first two weeks, and then cultured as airlifted culture in the following two weeks. Transmission electronmicroscope (TEM) was performed weekly, the AE-5 immunohistochemical staining, PAS staining and HE staining were performed at the end of fourth week.It took 2 weeks for the cultured limbal epithelium to cover the whole amniotic membrane. Immunocytochemical studies showed that cultured limbal epithelium was positive for keratin 3 and negative for PAS staining. HE staining showed that the morphologic characteristics of cultured limbal epithelium were similar to normal rabbit corneal epithelium. The adhesion complex formation didn't appear on 7th day and 14th day. Early stage of adhesion complex formation appeared on 21st day, and without significant change until 28th day.The morphologic characteristics of cultured limbal epithelium on amniotic membrane in vitro are similar to normal rabbit corneal epithelium. Only early stage of adhesion complex is formed as compare to the normal adhesion complex of rabbit.PubMed05426-304