Tissues maintained in vitro often undergo changes in the pattern of protein synthesis that result in the deposition of macromolecules quantitatively or qualitatively unrelated to those normally synthesized. In these preliminary studies, we modified published techniques to maintain adult and neonate rabbit corneas in vitro for 24 to 48 h. Measurements of corneal wet weight, and histologic and ultrastructural analyses were made to determine the success of maintaining rabbit corneas in culture. The results show that rabbit corneas can maintain normal corneal hydration and tissue structure for at least 48 h when incubated in Coon's modification of Ham's F12, 5% fetal or newborn calf serum, 2 mM glutamine, and 2% chondroitin sulfate or 2% 50 kDa dextran sulfate at 37 degrees C in a 5% CO2/air atmosphere. In addition, we confirmed previous observations that corneal explants must have a 1 to 2 mm rim of limbal sclera, and the organ placed in the culture dish with the epithelial side down to guard against damage and insure endothelial functioning. Normal ultrastructure of neonate rabbit cornea is also maintained when organ-cultured with these procedures. Moreover, neonate corneas continue to synthesize collagen in culture.