Ultraviolet Transmittance of Human Limbal Epithelial Cells Cultured on Human Amniotic Membranes

Abstract

Purpose: To evaluate ultraviolet (UV) A and B transmittance by human limbal epithelial cells cultured on human amniotic membranes. Methods: Human limbal epithelial cells were taken from the limbus of donor corneas and were cultured on human amniotic membranes with inactivated 3T3 fibroblasts for 2 to 4 weeks. Then, the cultured cells were examined histologically. Next, cells from different culture periods were irradiated with UV-A (365 nm) or UV-B (302 nm) at energy levels ranging from 50 to 800 μ W/cm², and UV transmittance was measured with a UV light meter. Results: Histological examination revealed a monolayer of corneal epithelial cells on the amniotic membrane after 2 weeks of culture, and a layer of 3–4 cells was formed after 4 weeks. Transmittance of UV-A and UV-B was highest by the amniotic membrane alone, followed in decreasing order by limbalepithelial cells cultured on amniotic membranes for 2 weeks, 3 weeks, and 4 weeks. Conclusions: These results indicate that UV absorbance increases in proportion to the number of limbal epithelial cell layers in cultures on amniotic membranes. Limbal epithelial cells may need to be cultured until 3–4 layers are formed in order to prevent ocular damage by UV light after transplantation.

KEYWORDS:

• amniotic membrane
• cell culture
• limbal epithelial cell
• ultraviolet