The effect of micropores in the surface of temperature-responsive culture inserts on the fabrication of transplantable canine oral mucosal epithelial cell sheets.

Primary canine oral mucosal epithelial cells were cultured on temperature-responsive dishes and cell culture inserts to fabricate transplantable epithelial cell sheets. When 3T3 feeder layers and fetal bovine serum were eliminated from dish culture, the harvested cell sheets became significantly more fragile. In contrast, when epithelial cells were cultured on inserts having submicron-scale pores, cell sheet fragility was eliminated. Keratin expression profiles showed no differences among the harvested cell sheets, but the expression of p63, a putative stem/progenitor marker, was strongly dependent on the presence of 3T3 feeder layers and serum. These results suggest that the maintenance of stem/progenitor cells is influenced by the apical/basal supply of nutrients as well as culture supplements.

[1]  H Green,et al.  Treatment of skin ulcers with cultured epidermal allografts. , 1989, Journal of the American Academy of Dermatology.

[2]  Masayuki Yamato,et al.  Functional bioengineered corneal epithelial sheet grafts from corneal stem cells expanded ex vivo on a temperature-responsive cell culture surface , 2004, Transplantation.

[3]  T. Okano,et al.  Corneal reconstruction with tissue-engineered cell sheets composed of autologous oral mucosal epithelium. , 2004, The New England journal of medicine.

[4]  Michele De Luca,et al.  Long-term restoration of damaged corneal surfaces with autologous cultivated corneal epithelium , 1997, The Lancet.

[5]  H. Green,et al.  Seria cultivation of strains of human epidemal keratinocytes: the formation keratinizin colonies from single cell is , 1975, Cell.

[6]  Benjamin Geiger,et al.  The catalog of human cytokeratins: Patterns of expression in normal epithelia, tumors and cultured cells , 1982, Cell.

[7]  C. Compton,et al.  Cultured Epithelial Autografts for Giant Congenital Nevi , 1989, Plastic and reconstructive surgery.

[8]  Masayuki Yamato,et al.  Transplantation of tissue-engineered epithelial cell sheets after excimer laser photoablation reduces postoperative corneal haze. , 2006, Investigative ophthalmology & visual science.

[9]  T. Okano,et al.  Thermo‐responsive polymeric surfaces; control of attachment and detachment of cultured cells , 1990 .

[10]  Masayuki Yamato,et al.  Ocular surface reconstruction using autologous rabbit oral mucosal epithelial sheets fabricated ex vivo on a temperature-responsive culture surface. , 2005, Investigative ophthalmology & visual science.

[11]  Amy Li,et al.  Location and phenotype of human adult keratinocyte stem cells of the skin. , 2004, Differentiation; research in biological diversity.

[12]  F. Gage,et al.  Human embryonic stem cells express an immunogenic nonhuman sialic acid , 2005, Nature Medicine.

[13]  G. Pellegrini.  Changing the cell source in cell therapy? , 2004, The New England journal of medicine.

[14]  A. Yang,et al.  p63, a p53 homolog at 3q27-29, encodes multiple products with transactivating, death-inducing, and dominant-negative activities. , 1998, Molecular cell.

[15]  T. Okano,et al.  Creation of designed shape cell sheets that are noninvasively harvested and moved onto another surface. , 2000, Biomacromolecules.

[16]  Masayuki Yamato,et al.  Cell-Sheet Engineering Using Intelligent Surfaces , 2005 .

[17]  H Green,et al.  Growth of cultured human epidermal cells into multiple epithelia suitable for grafting. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[18]  B. Doğan,et al.  Keratin 15 expression in stratified epithelia: downregulation in activated keratinocytes. , 1999, The Journal of investigative dermatology.

[19]  C. Compton,et al.  Permanent Coverage of Large Burn Wounds with Autologous Cultured Human Epithelium , 1984 .

[20]  Mitsuo Umezu,et al.  Fabrication of Pulsatile Cardiac Tissue Grafts Using a Novel 3-Dimensional Cell Sheet Manipulation Technique and Temperature-Responsive Cell Culture Surfaces , 2002, Circulation research.

[21]  T. Okano,et al.  Decrease in culture temperature releases monolayer endothelial cell sheets together with deposited fibronectin matrix from temperature-responsive culture surfaces. , 1999, Journal of biomedical materials research.

[22]  T. Okano,et al.  Thermo-responsive culture dishes allow the intact harvest of multilayered keratinocyte sheets without dispase by reducing temperature. , 2001, Tissue engineering.

[23]  G. Pellegrini,et al.  Autologous fibrin-cultured limbal stem cells permanently restore the corneal surface of patients with total limbal stem cell deficiency. , 2001, Transplantation.