June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Development of a Human Cultivated Nasal Mucosal Epithelial Cell Sheet for the Treatment of Severe Ocular Surface Diseases
Masakazu Kobayashi; Takahiro Nakamura; Makoto Yasuda; Yuiko Hata; Shoki Okura; Naoki Okumura; Noriko Koizumi; Hisao Yasu; Shigeru Kinoshita
Author Affiliations & Notes
  • Masakazu Kobayashi
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
  • Takahiro Nakamura
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Makoto Yasuda
    Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Yuiko Hata
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Shoki Okura
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
  • Naoki Okumura
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Noriko Koizumi
    Biomedical Engineering, Doshisha University, Kyotanabe, Japan
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Hisao Yasu
    Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Shigeru Kinoshita
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Footnotes
    Commercial Relationships Masakazu Kobayashi, None; Takahiro Nakamura, None; Makoto Yasuda, None; Yuiko Hata, None; Shoki Okura, None; Naoki Okumura, None; Noriko Koizumi, None; Hisao Yasu, None; Shigeru Kinoshita, Senju Pharmaceutical Co (P), Santen Pharmaceutical Co (P), Otsuka Pharmaceutical Co (C), Alcon (R), AMO (R), HOYA (R)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2561. doi:
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      Masakazu Kobayashi, Takahiro Nakamura, Makoto Yasuda, Yuiko Hata, Shoki Okura, Naoki Okumura, Noriko Koizumi, Hisao Yasu, Shigeru Kinoshita; Development of a Human Cultivated Nasal Mucosal Epithelial Cell Sheet for the Treatment of Severe Ocular Surface Diseases. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2561.

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Abstract

Purpose: We previously reported the establishment of a surgical method for cultivated corneal and oral mucosal epithelial cell-sheet transplantation for the treatment of severe ocular surface diseases. However, patients with severe dry eye can show a complete loss of goblet cells, resulting in those patients falling outside of the surgical indications. The purpose of this present study was to evaluate a new surgical method of autologous cultivated nasal mucosal epithelial cell-sheet transplantation that supplies goblet cells to the ocular surface for the treatment of severe ocular surface diseases.

Methods: After obtaining approval from the Institutional Review Board for Human Studies of Kyoto Prefectural University of Medicine, Kyoto, Japan, human nasal mucosa was cultivated for 2 weeks on a denuded amniotic membrane (AM) carrier with mesenchymal stem-cell co-culture and airlifting. The cultivated epithelium was examined by light microscopy, real-time polymerase chain reaction (PCR), and immunohistochemistry for several keratins and secretory mucins.

Results: A confluent primary culture of human nasal mucosal epithelial cells was established on AM after 7 days. After 2 weeks of culture, the human cultivated nasal epithelial sheet had 2-4 layers of stratified well-differentiated cells. Immunohistochemistry confirmed the presence of keratins 3, 4, 12, and13 in the cultivated nasal mucosal epithelial cells. Real-time PCR showed that the expression level of Muc5ac in the cultivated nasal mucosal epithelial cells was clearly elevated as compared with cultivated corneal and oral epithelial sheets.

Conclusions: The findings of this study show that a cultivated nasal epithelial cell sheet containing goblet cells was successfully generated, thus indicating that autologous cultivated nasal epithelial sheet transplantation can be used for treating severe ocular surface diseases.

Keywords: 482 cornea: epithelium • 474 conjunctiva • 485 cornea: surface mucins  
© 2013, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2015 Association for Research in Vision and Ophthalmology.
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