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Donor corneas can be preserved for ≤4 weeks in organ culture (31°C) by using modified minimal essential medium (MEM). About one fifth of them have to be discarded, however, as disintegration of the endothelial cell monolayer— enhanced polymegethism, cell loss—occurs. The objective of this study was to investigate whether addition of insulin, dextran, and epidermal growth factor (EGF) makes corneal endothelial cells more viable, stable, and homogeneous.Sixteen paired human donor corneas were cocultured in media supplemented with EGF and insulin-like growth factor (IGF) or in conventional modified MEM for 4 weeks. Endothelial parameters were evaluated at the outset and at days 7, 14, 21, and 28 of culture by using an automated digital image-analysis system.No significant differences were observed in the first 2 weeks of culture. Beginning with day 14, however, stabilization of endothelial cell patterns was evident for corneas cultured in supplemented culture media.Our data indicate that the addition of growth factors to culture media might increase the percentage of corneas available for transplantation and would also allow a significantly longer period of preservation.