September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Barrier function of cultured autologous oral mucosal epithelial cell sheets
Author Affiliations & Notes
  • Fawzia Bardag-Gorce
    Medicine, Los Angeles Biomedical Research Institute, Torrance, California, United States
  • Joan Oliva
    Medicine, Los Angeles Biomedical Research Institute, Torrance, California, United States
  • Andrew Makalinao
    Medicine, Los Angeles Biomedical Research Institute, Torrance, California, United States
  • Imara Meepe
    Medicine, Los Angeles Biomedical Research Institute, Torrance, California, United States
  • Julio Garcia
    Medicine, Los Angeles Biomedical Research Institute, Torrance, California, United States
  • Amanda M. Laporte
    Medicine, Los Angeles Biomedical Research Institute, Torrance, California, United States
  • Robert Niihara
    Medicine, Los Angeles Biomedical Research Institute, Torrance, California, United States
  • Hyo Jung Ha
    Medicine, Los Angeles Biomedical Research Institute, Torrance, California, United States
  • Richard H. Hoft
    Ophthalmology, Los Angeles Biomedical Research Institute, Torrance, California, United States
  • Alan L. Felsenfeld
    Oral and Maxillofacial Surgery, Harbor UCLA MEdical Center, Torrance, California, United States
  • Yutaka Niihara
    Medicine, Los Angeles Biomedical Research Institute, Torrance, California, United States
  • Footnotes
    Commercial Relationships   Fawzia Bardag-Gorce, None; Joan Oliva, None; Andrew Makalinao, None; Imara Meepe, None; Julio Garcia, None; Amanda Laporte, None; Robert Niihara, None; Hyo Jung Ha, None; Richard Hoft, None; Alan Felsenfeld, None; Yutaka Niihara, Emmaus Life Sciences (F)
  • Footnotes
    Support  Supported by CellSeed Inc., and Emmaus Life Sciences Inc.
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 895. doi:
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    • Get Citation

      Fawzia Bardag-Gorce, Joan Oliva, Andrew Makalinao, Imara Meepe, Julio Garcia, Amanda M. Laporte, Robert Niihara, Hyo Jung Ha, Richard H. Hoft, Alan L. Felsenfeld, Yutaka Niihara; Barrier function of cultured autologous oral mucosal epithelial cell sheets. Invest. Ophthalmol. Vis. Sci. 2016;57(12):895.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Cultured autologous oral mucosal epithelial cell sheet (CAOMECS) has been successfully used to regenerate corneal epithelium with limbal stem cell deficiency (LSCD). However, it is uncertain if corneal epithelium derived from CAOMECS has most of the barrier functions observed in normal corneal epithelium. Barrier functions include a barrier to conjunctivalization of the cornea, a barrier to external toxic and infectious agents, and barriers to stromal edema or dehydration. The present study reports the barrieral characterization of carrier free human CAOMECS (hCAOMECS).

Methods : Following informed consent procedures, a donated small biopsies was used to engineer hCAOMECS. Temperature responsive culture ware was used to harvest hCAOMECS with preserved extra cellular matrix, which permits a quick adherence to corneal stroma, and plays the role of conjunctival barrier (previously reported in our rabbit study (Bardag-Gorce et al, Ocul Surf. 2015)).

Results : Immuno-histological analysis showed that hCAOMECS is a multilayered epithelial tissue-like cell sheet with apico-basal polarity evidenced by basal expression p63 and Ki67, and apical expression of K4/K13. hCAOMECS also expressed pro-angiogenic factors, such as matrix metalloproteinase (MMP-3) in balance with the anti-angiogenic tissue inhibitors of metalloproteinases (TIMP-3), which suggest that hCAOMECS has characteristics of a normal corneal epithelium. hCAOMECS epithelial markers were also investigated; the results showed positive expression of E-cadherin, alpha-catenin, and beta-catenin at the cell borders. While beta-catenin levels were similar, E-cadherin and Occludin levels were found significantly higher in hCAOMECS compared to cultured monolayer human corneal epithelia cells, indicating functional adherens and tight junctions. These results were repeatedly obtained in our protocols of engineering rabbits or human CAOMECS. E-cadherin has been used to examine potential cell tumorigenicity and is essential for organization of the cytoskeleton and junctional complexes. In our studies, we use E-cadherin expression as an indicator of a functional epithelial barrier because it solidifies intercellular adhesion, recruits catenins to cell borders, and recruits actin filaments to increase barrier function.

Conclusions : hCAOMECS has high expression of epithelial markers necessary for normal corneal epithelial barrier function.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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