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 2001-2002

Development and Use of an Immortalized Human Goblet Cell Line for the Use in Ocular Toxicity Testing

Darlene Dartt, PhD
The Schepens Eye Research Institute, Harvard University, Boston, Massachusetts

The conjunctival epithelium along with the cornea make up the ocular surface. The conjunctiva has long been used as a component of the Draize test. Mucin secretion from conjunctival tissue has been one of the endpoints measured by this test. The Draize test has been the accepted method by which a variety of agents have been evaluated with respect to their ability to cause irritation and injury to the ocular surface. Unfortunately the inhumane treatment of animals during this test procedure has often been overlooked. To replace the Draize test, many laboratories have used a variety of cell lines and primary cultures of corneal epithelial cells and conjunctival squamous cells. No one, however, has been able to culture conjunctival goblet cells. The long term goal of our proposal is to develop a cell line of human conjunctival goblet cells with an extended life span which will be able to be used for ocular irritancy testing. We hope to accomplish this goal by fulfilling the following specific aims:

  1. Develop a cell line witn an extended life span by telomerase transfection of primary cultures of human goblet cells;
  2. Characterize the new cell line by using established morphological, histochernical, immunocytochemical and biochemical markers known to be associated with goblet cells; and
  3. Determine the effects of neurotransmitters and growth factors on mucin and protein secretion from this cell line and the effect of ocular irritants on these secretions.

The development of a human goblet cell line with an expanded lifespan that secrete mucin and other antibacterial proteins is a useful model that could be potentially use in place of the Draize test.