Isolation of oral epithelial progenitors using collagen IV
Article first published online: 28 JUN 2008
© 2008 The Authors. Journal compilation © 2008 Blackwell Munksgaard
Volume 14, Issue 5, pages 413–418, July 2008
How to Cite
Igarashi, T., Shimmura, S., Yoshida, S., Tonogi, M., Shinozaki, N. and Yamane, G.-y. (2008), Isolation of oral epithelial progenitors using collagen IV. Oral Diseases, 14: 413–418. doi: 10.1111/j.1601-0825.2007.01390.x
- Issue published online: 28 JUN 2008
- Article first published online: 28 JUN 2008
- Received 9 February 2007; revised 9 February 2007; accepted 13 February 2007
- oral epithelial cells;
- progenitor cells;
- type IV collagen;
- colony forming efficiency assay;
Objective: Although oral mucosal epithelial stem cells are thought to reside in the basal layer, such cells have not yet been isolated. We isolated a population of rabbit oral epithelial progenitor cells containing putative stem cells.
Materials and methods: Epithelial cells harvested from rabbit buccal mucosa were allowed to adhere to dishes coated with collagen IV for periods ranging from 10 min to 16 h. The properties of individual cell populations were evaluated using BrdU, Ki-67, integrin β1, integrin α6 and keratin 13 using colony forming efficiency (CFE).
Results: Cells that adhered to collagen IV-coated dishes within 10 min were enriched about sixfold in terms of BrdU incorporation, Ki-67, integrin α6 and integrin β1 were strongly expressed. Interestingly, keratin 13 was faintly expressed. The CFE of rapidly adherent cells among oral epithelial cells was significant compared with other cell populations.
Conclusions: These results suggested that rabbit oral epithelial cells could be isolated by depending on adhesiveness to collagen IV, especially when segregated according to progenitor cell properties. Putative progenitor cells with stem cell properties were most effectively harvested within 10 min. Our separation procedure should be a useful tool with which to isolate epithelial stem cells for regenerative medicine.