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

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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  
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