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Abstract

Purpose Ocular surface reconstruction with cultivated oral mucosal epithelial transplantation technique has potential in the treatment of patients with severe ocular surface injuries. Currently, this technique is mainly based on utilization of xenogenic/allogenic components such as murine feeders, serum and amniotic membrane. The use of animal-derived materials possesses risk of pathogen transmission, immune reactions and graft rejection.

Methods The formation of stratified sheets by human oral mucosal epithelial cells under serum-free culture environment both in the absence and presence of fibroblast-conditioned culture medium and elevated epidermal growth factor concentration was examined. The integrity of the epithelium was measured by transepithelial electrical resistance. The tissue-engineered cell sheets were also studied for histology and immunohistochemical markers for epithelial keratins (K), cell proliferation and cell adhesion.

Results In all examined culture conditions, the cultivated oral epithelial cells formed a stratified tissue positive for keratins K3/12, K4, and K13. The tissue-engineered oral epithelia also expressed proliferation and progenitor markers Ki67 and p63 in the basal layer. The cultures presented expression of tight junction proteins ZO-1 and occludin and high transepithelial electrical resistance values.

Conclusion Tight multi-layered epithelium with proliferative potential can be produced from human oral mucosal epithelial cells under defined culture conditions and without the use of serum.