[The interactions of stromal-epithelial in a model of co-culture in the rabbit cornea].

OBJECTIVE

The purpose of the study is to produce a reconstructed cornea including epithelia and stroma by tissue engineering. The reconstructed tissue may provide a physiologic model for the investigations of interaction between corneal epithelial cells and keratocytes.

METHODS

Epithelial cells and keratocytes were isolated from rabbit corneas and cultured on plastic substrates in vitro. The co-culture model was established using a special transwell in which two different cells were separated but were able to interact each other. Histological and immunohistological studies were performed to identify the cell types. Intercellular communication of both the cultured epithelial cells in pure and in co-culture with the keratocytes was studied by laser confocal scanning microscopy.

RESULTS

Population doubling time (PDT) was 3.45, 3.30, 2.11 and 2.32 d in pure corneal epithelial cells, co-culture epithelial cells, pure keratocytes and co-culture keratocytes respectively. The epithelial cells in co-culture grew quicker than those in pure (P < 0.01) and the stromal cells in co-culture grew slower than those in pure (P < 0.01). Intercellular communication of the cultured epithelial cells in co-culture were more than that in pure (U = 2.691, P < 0.05).

CONCLUSIONS

The co-culture model of epithelial cell and keratocyte is feasible. Under the co-culture system the responses of cell proliferation and intercellular communication are different between epithelial cells and keratocytes.