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Investigative Ophthalmology & Visual Science, Vol 34, 3442-3449, Copyright © 1993 by Association for Research in Vision and Ophthalmology

ARTICLES AND REPORTS

Requirement for vitamin A in long-term culture of human cornea

JA Anderson, NR Richard, ME Rock and PS Binder
National Vision Research Institute, Sharp Cabrillo Hospital, San Diego, California 92103.

PURPOSE. To develop an in vitro model for the study of vitamin A functions in the cornea by determining the morphologic responses to its absence and to the addition of varying concentrations in long-term (21- day) human organ culture. METHODS. Donor corneas were cultured for 21 days in an air-liquid environment designed to maintain normal corneal morphology in long-term culture. Corneas were graded by light microscopy and were further analyzed by transmission electron microscopy. RESULTS. Epithelium of human corneas cultured in the absence of vitamin A for 21 days showed characteristics of vitamin A deficiency: multilayering of squamous cells, bundling of tonofilaments to form keratofibrils, increased desmosomes, and loss of microplicae on the surface cells. The cells responded to the addition of vitamin A in concentrations ranging from 5 x 10(-9) to 5 x 10(-6) M. At the lowest concentration, cell differentiation and morphology appeared normal. Cells developed secretory morphology at higher concentrations of vitamin A, with intracellular vesicles, basement membrane overproduction, and reduced cell-cell and cell-substrate attachments. Keratocytes and endothelial cells showed increased intracellular vacuoles in the presence of higher concentrations of vitamin A. CONCLUSIONS. Cultured adult human corneas showed primary, dose-related responses to vitamin A in the absence of inflammatory, neuronal, or humoral factors. The epithelium was most sensitive to the absence of vitamin A and showed a dose-related response to additional vitamin A. All corneal cell layers showed morphologic changes at the highest concentration of vitamin A used.


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L. Liu, D. Hartwig, S. Harloff, P. Herminghaus, T. Wedel, K. Kasper, and G. Geerling
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