Three-dimensional Culture Model of the Conjunctival Epithelium

ABSTRACT

PURPOSE: To reconstruct a cultured conjunctival equivalent that closely resembles normal conjunctival epithelium in three-dimensional culture systems. METHODS: Human conjunctival epithelial cells were cultured on dead de-epidermized dermis in the air-exposed state. After 2 weeks of culture, the sections were stained with hematoxylin and eosin. Immunohistochemical and electron microscopic studies were performed. The results were compared with those of normal conjunctiva and cultured eyelid skin equivalent. RESULTS: In the cultured conjunctival equivalent, nonkeratinizing stratified epithelium was formed similarly to normal conjunctival epithelium. Keratin 13 was expressed, but not keratin 10, in the cultured conjunctival equivalent, similarly to normal conjunctival epithelium. However, in the cultured eyelid skin equivalent, keratinizing stratified epithelium was formed. In addition, keratin 10 was expressed, but not keratin 13, contrary to those of the cultured conjunctival equivalent. In the cultured conjunctival equivalent, ultrastructurally, keratin intermediate filaments and desmosomes were found. In addition, microvilli were seen in the uppermost epithelial cells. CONCLUSIONS: These findings demonstrate that the cultured conjunctival equivalent resembles normal conjunctival epithelium morphologically, biochemically and ultrastructurally, thereby suggesting that the cultured conjunctival equivalent may have a great potential in the study of conjunctival epithelium.