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H.-J. Jin, J.-S. Choi, C.-K. Joo; Wnt4 Promotes Proliferation and Maintaining of Limbal Stem Cells (LSc). Invest. Ophthalmol. Vis. Sci. 2008;49(13):6075.
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In order to proliferate and maintain stemness of human limbal stem cells, we investigated using Wnt proteins.
Primary human limbal epithelial cells were gained from rime of donor corneas. The wnt transfected NIH 3T3 cells were used as feeder cell in this study. The limbal epithelial cells were co-cultured on MMC treated NIH 3T3 cells. The wnt expression were confirmed by expression of HA-tagging protein. After co-culture, we confirmed colony size and expression of limbal stem cell marker. P63, CD34 and connexin 43 were observed by western blot analysis.
Isolated limbal stem cells were co-cultured on NIH 3T3 cells with or without expression of wnt protein. In wnt treated limbal stem cells, the size of colonies were larger than it in non-treated limbal stem cells. The beta-catenin translocation was observed in wnt treated groups. And the CD34 were strongly expressed in wnt treated limbal stem cells. The p63 were expressed in both group. The connexin 43 and keratin 3, marker of differentiation, was not expressed in wnt treated group.
Wnt4 increased the proliferation maintaining of limbal stem cells in culture system. Our study suggest that wnt4 signaling can control proliferation and fate of the limbal stem cells.
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