Isolation of oral epithelial progenitors using collagen IV.
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.


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 beta1, integrin alpha6 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 alpha6 and integrin beta1 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.
Igarashi
T
T
Department of Oral Medicine, Oral and Maxillofacial Surgery, Tokyo Dental College, Chiba, Japan. igarasit@jcom.home.ne.jp
Shimmura
S
S
Yoshida
S
S
Tonogi
M
M
Shinozaki
N
N
Yamane
G-y
GY
D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D018929:Cell Culture Techniques; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000502:physiology*; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D039503:Integrin alpha6 / Q000378:metabolism; D019012:Integrin beta1 / Q000378:metabolism; D053535:Keratin-13 / Q000378:metabolism; D019394:Ki-67 Antigen / Q000378:metabolism; D009061:Mouth Mucosa / Q000166:cytology*; D011817:Rabbits; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
D024141:Collagen Type IV; D039503:Integrin alpha6; D019012:Integrin beta1; D053535:Keratin-13; D019394:Ki-67 Antigen