Abstract
Abstract: :
Purpose: To compare the growth rate of human limbal explant cell cultures established on human amniotic membrane with cell cultures established on laminin, hyaluronic acid, collagen I, collagen IV and fibronectin. Methods: Six human limbal explants were prepared from a single corneo – scleral rim. The cell cultures were established under identical cell culture conditions and grown within cell culture inserts on human amniotic membrane, hyaluronic acid, collagen I, collagen IV, fibronectin, laminin and co–cultured with mitomycin C inactivated 3t3 fibroblasts. The cell cultures were examined by light microscopy and serial digital photographs were obtained. The mean cell culture growth distance after twenty one days was calculated by novel computer software developed by the stem cell research group at Kingston University. Results: The computer software calculated mean growth distances after twenty one days were: amniotic membrane – 0.43 mm; Hyaluronic acid – 0.22 mm; Collagen IV – 0.24 mm; Laminin – 0.22 mm; Fibronectin – 0.09 mm and Collagen I – 0.14 mm. Conclusions: Amniotic membrane produced cell cultures with the highest mean cell culture growth distances. The release of intrinsic growth factors and the structure of the basement membrane may explain the observed results. Collagen IV, laminin and hyaluronic acid all produced similar growth distance values, however the mean growth distances were almost half that observed with amniotic membrane. The use of Collagen I and fibronectin produced very low growth distances and these were considered sub–optimal substrates for cell culture.
Keywords: cornea: epithelium • cornea: basic science • wound healing