May 2006
Volume 47, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2006
Investigation of Putative Stem/Progenitor Cells in Human Oral Mucosal Epithelial Cell Sheets Harvested From Temperature–Responsive Culture Surface
Author Affiliations & Notes
  • H. Sugiyama
    Department of Ophthalmology, Osaka University Medical School, Suita, Japan
  • M. Yamato
    Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan
  • Y. Hayashida
    Department of Ophthalmology, Osaka University Medical School, Suita, Japan
    Ocular Surface Research and Education Foundation, R&D Department, Tissue Tech, Inc., Miami, FL
  • T. Soma
    Department of Ophthalmology, Osaka University Medical School, Suita, Japan
  • R. Hayashi
    Department of Ophthalmology, Osaka University Medical School, Suita, Japan
  • H. Takayanagi
    Department of Ophthalmology, Osaka University Medical School, Suita, Japan
  • A. Kikuchi
    Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan
  • T. Okano
    Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan
  • Y. Tano
    Department of Ophthalmology, Osaka University Medical School, Suita, Japan
  • K. Nishida
    Department of Ophthalmology, Tohoku University School of Medicine, Sendai, Japan
  • Footnotes
    Commercial Relationships  H. Sugiyama, None; M. Yamato, CellSeed C, P; Y. Hayashida, None; T. Soma, None; R. Hayashi, None; H. Takayanagi, None; A. Kikuchi, CellSeed C, P; T. Okano, CellSeed C, P; Y. Tano, None; K. Nishida, CellSeed C, P.
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science May 2006, Vol.47, 3937. doi:
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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)

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Abstract

Purpose: : 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.

Methods: : 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.

Results: : 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.

Conclusions: : 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.

Keywords: cornea: epithelium • regeneration • clinical laboratory testing 
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