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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. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
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.
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.
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.
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.
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