Optimization of the Culture Conditions for Feline Corneal Endothelial Cell Cultures

Abstract

Purpose: : To evaluate the effect of various culture conditions and growth supplements on feline corneal endothelial cell (FCEC) cultures.

Methods: : FCEC were seeded on plastic or on gelatin-coated plates and cultured either with serum alone (0, 5, 10, 20%), with serum and epidermal growth factor (EGF; 0.5, 5, 25, 100 ng/ml) with serum and bovine pituitary extract (BPE; 25, 100 µg/ml) or a combination of both. Cell number, size and morphology were assessed on three different cell populations. Cell cultures were also maintained at confluence to observe the post-confluence changes in cell morphology.

Results: : Culturing FCEC on gelatin-coated plates greatly helped in the preservation of an endothelial morphology. EGF and BPE induced proliferation above basal level. The combination of gelatin-coating and supplementing the growth medium with 10% serum, EGF and BPE increased cell numbers above those achieved with serum alone while maintaining a normal endothelial morphology.

Conclusions: : Improvements have been made for the culture of FCEC. The final selected medium consistently allows the growth of a contact inhibited cell monolayer of small, polygonal-shaped cells.

Keywords: cornea: endothelium • cornea: basic science • growth factors/growth factor receptors