Perfusion culture promotes differentiation of oral keratinocytes in vitro.

ABSTRACT

In order to reconstruct the mucosal lining of the oral cavity tissue engineered autologous mucosa grafts could be of great benefit. In conventional stagnant cultures cells often tend to dedifferentiate. Perfusion culture has been demonstrated to reestablish differentiation in various epithelial cell types. Thus, in secondary cultures of human oral keratinocytes from ten patients conventional stagnant culture versus perfusion culture technique was compared. Proliferation and state of differentiation as expressed morphologically and immunohistochemically were assessed. After 14 days oral keratinocytes in a perfusion culture system tend to be further differentiated. They build up a thicker epithelium (3.4+/-1.0 vs. 2.4+/-0.4 layers), form microridges, express cytokeratins 1, 2, 10, 11, 19 but not 13 in all cells and cytokeratins 5, 6 particularly in cells attached to the carrier membrane. In contrast, oral keratinocytes in stagnant culture do not form microridges and rather express cytokeratins 13, 14, and 19 in mattering amounts.