Molecular Characterization of Explant Cultured Human Oral Mucosal Epithelial Cells

Abstract

To culture and characterize oral mucosal epithelial cells (OMEC) grown on de-epithelialized human amniotic membrane (HAM) to explore their suitability as autografts in patients with bilateral ocular surface disease (OSD) and limbal stem cell deficiency. Oral biopsy samples were obtained from 20 patients undergoing oral reconstructive surgery, with informed consent and Institutional Ethics Committee approval. Morphologic studies, transmission electron microscopy (TEM), reverse transcriptase (RT) PCR and immunocytochemistry were used to characterize the OMEC. Morphologic studies and TEM revealed a confluent sheet of proliferating, stratified oral epithelial cells connected to each other by desmosomes, containing intracellular cytokeratins and abundant mucin granules. These characteristics were further corroborated and elucidated by RT-PCR and immunocytochemistry. The presence of markers of differentiated, stratified epithelial cells (cytokeratin K3, K4, K13, and connexin 43), progenitor stem cell cell markers (p63, p75, β1-integrin/CD29, and ABCG2), and a variety of predominantly membrane-bound and a few gel-forming mucins (MUC 1, 5B, 6, 13, 15, and 16) was established. Cultured OMEC have the potential to act as autografts for ocular surface reconstruction in patients with bilateral ocular surface disease and can prove to be particularly beneficial to ameliorate the mucin deficiency state in dry eye associated with OSD.