Quantification of corneal organ culture migration: central and peripheral epithelium

This report outlines a method for rapid quantification of corneal epithelial cell movement in a block organ culture system designed to model a penetrating or perforating wound of the cornea. Rabbit corneas were secured over a plastic conformer with a chalazion clamp. Two-millimeter, full-thickness corneal specimens were sampled from central and peripheral cornea and placed in tissue culture. The aim of the procedure was to produce full-thickness corneal specimens from well-defined geographic areas of the cornea of reproducible size and shape without undue trauma to the epithelium. The instruments used for measurements were simple, allowing for accumulation of a large number of observations which could be subjected to rigorous statistical evaluation. This method was used to determine if there was a detectable difference in the rate of epithelial cell movement over stromal cut surface of specimens from the periphery of the cornea, versus the rate over specimens from the central area of the cornea. In this block organ culture system the peripheral epithelial cells migrated more rapidly than samples of central epithelial cells (P less than 0.05). This study shows a simple, rapid method for producing and evaluating block corneal organ cultures and shows that the site of specimen sampling must be controlled in corneal epithelial cell studies employing block organ culture. This method will be useful for in vitro screening of novel pharmacologic agents which have the potential for influencing corneal wound healing prior to evaluating these agents in vivo.