Towards a defined, serum- and feeder-free culture of stratified human oral mucosal epithelium for ocular surface reconstruction

Tanja Ilmarinen, Juhana Laine, Kati Juuti-Uusitalo, Jura Numminen, Riitta Seppänen-Suuronen, Hannu Uusitalo, Heli Skottman

Research output: Contribution to journalArticleScientificpeer-review

19 Citations (Scopus)


Purpose: Ocular surface reconstruction with cultivated oral mucosal epithelial transplantation technique is a viable treatment option for severe ocular surface injuries and diseases with limbal stem cell deficiency. Currently, this technique is based on utilization of xenogenic, allogenic or undefined components such as murine 3T3 feeders, serum and amniotic membrane. In this study, we aimed to find a more defined culture method to generate stratified human oral mucosal epithelium. Methods: In this study, we have examined the formation of stratified cell sheets from 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 (EGF) concentration. Results: In all examined culture conditions, the cultivated oral epithelial cells formed a stratified tissue, which was 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 of the cell sheets, suggesting that the epithelia still had regenerative capacity. The cultures presented expression of tight junction proteins ZO-1 and occludin and high transepithelial electrical resistance values. Conclusion: In this culture method, we have been able to produce stratified cell sheets successfully without serum, conditioning of the medium or increased EGF concentration. We provide a novel protocol to produce tight multi-layered epithelium with proliferative potential, which can be easily adapted for cultivated oral mucosal epithelial transplantation.

Original languageEnglish
Pages (from-to)744-750
Number of pages7
JournalActa Ophthalmologica
Issue number8
Publication statusPublished - 2013
Publication typeA1 Journal article-refereed


  • corneal epithelium
  • defined
  • feeder-free
  • ocular surface reconstruction
  • oral mucosal epithelium
  • serum-free

Publication forum classification

  • Publication forum level 2


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