Abstract
Purpose::
Monkey corneal endothelial cells (MCECs) are thought not to possess replicative capacity in vivo. Cells cultured in vitro, on the other hand, do show a capacity to replicate. To help reveal the mechanisms regulating corneal endothelial cell proliferation, the expression of cell cycle-associated proteins were determined in MCECs in vivo and in vitro.
Methods::
Whole-mount fixed preparations of cynomolgus monkey corneas and confluent cultivated cynomolgus MCECs passaged on collagen type 4 coated plates were prepared for immunohistochemical analysis. Confocal microscopy was used to study the expression of the following cell cycle-associated proteins: Rb, cyclin A, p16, p21, p53, and phosphorilated p53.
Results::
MCECs in vivo showed nuclear staining for Rb, p16, and p21, but no staining for cyclin A, p53, or phosphorilated p53. Meanwhile, cultivated MCECs showed nuclear staining for Rb, p16, p21 and p53, but no staining of phosphorilated p53. In occasional cultivated cells the nucleus stained positively for cyclin A.
Conclusions::
We have demonstrated a different pattern of cell cycle-associated molecular expression between in vivo and in vitro MCECs. This information goes some way towards helping us understand the molecular mechanisms regulating the cell cycle in the corneal endothelium.
Keywords: cornea: endothelium • cornea: basic science • immunohistochemistry