Combined effect of extracellular matrices and growth factors on bovine corneal endothelial cells in culture

Purpose: First, to confirm that corneal endothelial cells in the confluent state have the capability to form cellular covering. Second, to establish a method to study the combined effect of extracellular matrices (ECMs) and g rowth factors on the biological response in corneal endothelial cells in cu lture. Methods: Bovine corneal endothelial cells were cultured inside a cylinder s et on a plastic dish. They formed a confluent cell nest on the dish coated with type I or type IV collagen, laminin, or fibronectin. After the removal of the cylinder, hepatocyte growth factor (HGF), epidermal growth factor, transforming growth factor-alpha or transforming growth factor-beta (1) was added to the cultures. Each confluent cell nest enlarged outward, and its increased area size was measured. Cellular response in the nest, including cellular proliferation, was analyzed. Results: The size of the increased area of the culture on type IV collagen plus HGF was the largest of all the combinations of ECMs and growth factors . The responses of component cells in the increased area consisted of cellu lar hypertrophy, proliferation, migration and giant cell formation. The tre atment with type IV collagen plus HGF clearly promoted all the above respon ses. Conclusions: The biological response of corneal endothelial cells was regul ated by ECMs and growth factors. (C) 2001 Japanese Ophthalmological Society .