Corneal endothelium is the single-cell layer that forms a physical barrier between the corneal stroma and aqueous humour. The barrier and ionic 'pump' functions of corneal endothelium help regulate stromal hydration. Loss of endothelial cells due to increasing age, trauma, disease, dystrophy, or previous corneal transplants can reduce the density of endothelial cells to a critical point below which the stroma becomes edematous and visual acuity is lost. Throughout life, division of endothelial cells either does not occur or occurs at a rate too slow to adequately replace dead cells. Thus, the major means of repairing the monolayer is by cell migration and/or enlargement. The basis for the lack of endothelial cell proliferation is not yet fully understood, although it is clear that cells do retain proliferative capacity. Previous studies from this laboratory have identified certain environmental conditions that may be responsible for maintaining these cells in a non-replicative state in vivo. In addition, corneal endothelial cells exhibit intrinsic, age-related differences in relative proliferative capacity. The studies described below provide evidence to support the hypothesis that, with age, an increasing number of HCEC enter a replicative senescence-like state in which they become increasingly refractive to mitogenic stimulation. This decreasing sensitivity to mitogens appears to be mediated, at least in part, by age-dependent alterations in the relative expression and activity of the cyclin-dependent kinase inhibitors, p27KIP1, p16INK4A, and p21CIP1.