| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2011: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2010: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2009: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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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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