Abstract
Purpose: :
We recently reported that N-cadherin-expressing conjunctival epithelial cells have strong stem-cell properties. The purpose of this present study was to construct three-dimensionally cultured epithelial tissue from N-cadherin-expressing conjunctival epithelial cells and investigate the histochemistry and expression profile of the tissue.
Methods: :
From single cell suspension of conjunctival epithelial cells prepared by the use of dispase and trypsin, N-cadherin-positive conjunctival epithelial cells were magnetically separated and then seeded on an atelocollagen gel containing mitomycin C-treated 3T3 cells. After the cells had reached 100% confluence, they were cultured under an airlifted condition. For the epithelial tissue, expression analysis against various types of undifferentiated-state, differentiated-state, and cell adhesion related markers was performed.
Results: :
Conjunctival epithelium cultured from the N-cadherin-positive cells exhibited a stratified squamous epithelium. In the three-dimensionally cultured conjunctival epithelial tissue, N-cadherin, p63, and cytokeratin 15 as undifferentiated-state markers were found to be expressed in the basal layer, cytokeratin 4 was found to be expressed in the suprabasal to upper layer, and cytokeratin 13 was found to be expressed in all layers. ZO-1 and desmoplakin, both known to be cell adhesion-related markers, were also expressed in the tissue.
Conclusions: :
Findings derived from the obtained data for the expression pattern of the undifferentiated-state, the differentiated-state, and the cell adhesion related markers show that N-cadherin-expressing conjunctival epithelial cells have a capacity to maintain stem cell population as well as to create moderately and terminally differentiated cell populations in an artificially configured organotypic culture system.
Keywords: conjunctiva • regeneration • cell adhesions/cell junctions