The Center for Alternatives to Animal Testing is an academic center affiliated with the Division of Toxicological Sciences in the Department of Environmental Health Sciences of the Johns Hopkins University Bloomberg School of Public Health.

 

Johns Hopkins School of Public Health

Research Grants 2000-2001

In Vitro Irritancy Test Using Telomerase Transfected Human Corneal Cells

James V. Jester, PhD
The University of Texas Southwestern, Dallas, Texas

Eye irritation testing is recognized as important in determining the safety of consumer products where manufacture or use may lead to accidental exposure and damage to the eye. Irritancy testing as currently performed, however, requires the use of live animals for which there are no recognized alternative replacement tests. The long-range goal of our work is to first develop and then validate an alternative, replacement test using a human tissue culture model that reconstructs the anterior, exposed portion of the human eye. A critical first step will be the generation of extended life-span human cells from the anterior part of the eye or cornea that show structural and functional characteristics similar if not identical to normal cells. We are currently inserting into corneal cells a gene encoding the enzyme telomerase that controls the number of times a cell divides, greatly extending if not indefinitely, the life the cell while maintaining the normal cellular characteristics unlike cells that are immortalized using various cancer genes. During the next year we will continue to clone human corneal epithelial, keratinocyte and endothelial extended life-span cell lines. We will also begin to establish and characterize various 3-dimensional corneal tissue constructs using these cells and determine how closely these constructs resemble normal corneal tissue. In the last year of this project we intend to test the response of these constructs to known ocular irritants.