Ex vivo Properties of Cornea Reconstructed with Cultured Human Corneal Endothelial Cells

Abstract

Abstract: : Purpose: To examine ex vivo properties of corneas reconstructed with cultured human corneal endothelial cells (HCEC) and human corneal stroma. Methods: Primary cultures of HCEC were established from explants of the endothelial cell layer and propagated on culture dishes coated with bovine corneal endothelial extracellular matrix. Cell suspension of HCEC at the fifth passage was transferred onto the human corneal stroma deprived of endothelial cells and the corneas were gently centrifuged to enhance the cell attachment. The reconstructed corneas were stained with alizarin red and trypan blue and the cell density was calculated. The reconstructed corneas underwent histological examination with light and electron microscope. The pump function of the reconstructed corneas was measured using Ussing chamber. Results: The mean endothelial cell densities of four reconstructed corneas was 2,380 ± 264 cells/mm² (mean ± S.D.). HCEC on the reconstructed corneas has similar morphology to HCEC in vivo. The parameters of the pump function of the reconstructed corneas were 55-75 % of those of normal corneas. Conclusions: HCEC on the reconstructed corneas has similar morphology and cellular density to HCEC ex vivo whereas the pump function of the reconstructed corneas was relatively low. Novel methods or technology enhancing the function of the reconstructed cornea are needed.

Keywords: cornea: endothelium • cornea: basic science