2011 Fiscal Year Final Research Report
Role of N-cadherin in the niche of corneal limbal ephthelium
| Project/Area Number |
21791711
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Ophthalmology
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| Research Institution | Keio University |
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Principal Investigator |
HIGA Kazunari 慶應義塾大学, 医学部, 共同研究員 (60398782)
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| Project Period (FY) |
2009 – 2011
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| Keywords | N-cadherin / ニッシェ / 角膜上皮前駆細胞 / siRNA / インターラクション |
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| Research Abstract |
To demonstrate the role of N-cadherin(N-cad), we observed the limbal basal epithelial layer by historological analysis and maintaining the progenitor status of primary human limbal epithelial cells in vitro. AQP1 positive cells(AQP1+) were observed as non-epithelial cells in the subepithelial stroma. When we made a thorough search of limbal basal cells by confocal microscopy, AQP1+were observed in the proximity of N-cad, K15 and p63 positive limbal basal epithelial cells. Electron microscope revealed the stromal cells penetrated the epithelial basal membrane and formed calcium-dependent cellular adhesions with N-cad+limbal basal epithelial cells. Furthermore, colonies using N-cad^<low> 3T3 cells were significantly smaller than mock-transfected cells. We then used a duplex-feeder model using two layers of either 3T3 or N-cad^<low> 3T3 cells to produce stratified epithelial sheets. Only N-cad+3T3 cells produced epithelial sheets with basal K15/suprabasal K12 expression observed in limbal tissue. AQP1+cells immediately beneath the epithelial basement membrane may be mesenchymal niche cells that directly interact with N-cad+limbal basal epithelial progenitor cells. N-cad plays a pivotal role in the maintenance of the progenitor phenotype in cultured limbal epithelial cells.
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