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S. Shimmura, H. Miyashita, S. Yoshida, J. Shimazaki, K. Tsubota; The Use of Keratocyte Progenitors as Feeder Cells for Epithelial Sheet Cultivation . Invest. Ophthalmol. Vis. Sci. 2006;47(13):2713.
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© ARVO (1962-2015); The Authors (2016-present)
To compare feeder cells prepared from mouse corneal stromal progenitors (COPs) with 3T3 fibroblasts.
Mouse stormal progenitor cells (COPs) were cultured and passaged as spheres in serum–free medium. COPs were then seeded onto plastic dishes with serum and treated with MMC to arrest growth. MMC–treated NIH/3T3 cells were used as control. Prolierfation of primary human limbal epithelium was analyzed by colony forming efficiency (CFE). Stratified epithelial sheets were prepared by air–lift cultures, which were compared by immunochytochemistry against K3, Connexin 43 (Cx43), integrin ß1 and p63.
Isolated colonies of primary human limbal epithelial cells were observed in both COP feeder and 3T3 feeder groups. CFE was significantly lower with COP feeders cells (p<0.05), however, both feeders produced stratified eptihelial sheets expressing similar levels of K3, Cx43, integrin ß1 and p63.
Feeder cells prepared from COPs can be used as an alternative to 3T3 feeder cells for preparing stratified epithelial sheets. Since COPs can be exanded indefinitely in vitro, the isolation of human COPs may allow cell sheet cultivation without the use of xenogeneic cells.
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