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H. Sugiyama, M. Yamato, Y. Hayashida, T. Soma, R. Hayashi, H. Takayanagi, A. Kikuchi, T. Okano, Y. Tano, K. Nishida; Investigation of Putative Stem/Progenitor Cells in Human Oral Mucosal Epithelial Cell Sheets Harvested From Temperature–Responsive Culture Surface . Invest. Ophthalmol. Vis. Sci. 2006;47(13):3937.
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© ARVO (1962-2015); The Authors (2016-present)
We previously reported a novel method for transplanting tissue–engineered cell sheets of oral mucosal epithelium to ocular surfaces using a temperature–responsive cell culture surface. We now demonstrate the presence of viable stem/progenitor cells in these human oral mucosal epithelial cell sheets.
Stem/progenitor cell populations derived from primary epithelial cells obtained from human oral mucosal tissue and from secondary cells collected by dissociation of harvested cell sheets were examined by colony–forming assay. The expression patterns of integrin alpha–6 and p63, which are known marker of putative epithelial stem/progenitor cells in human oral mucosal tissue and the harvested cell sheets were examined by immunofluorscence.
Colony–forming efficiency was 1.70 ± 0.21% (mean ± SE, n = 19) for primary cell sources subjected to cell sheet fabrication, and 1.26 ± 0.29% (mean ± SE, n = 19) for the total cells of the harvested cell sheets. Considering cell proliferation and resultant in total cell number, the number of colony–forming progenitor cells in the harvested cell sheets increased 285.9% from the primary seeded stem/progenitor cells. Immunofluorescence showed that integrin alpha–6 and p63 were expressed only in the basal layer of oral mucosal tissue and cell sheets.
These results are consistent with retention of stem/progenitor cells in the basal layer of stratified human oral mucosal epithelial cell sheets fabricated on temperature–responsive culture dishes. This maintenance is of importance for long–term graft survival.
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