Figure 1. Gross pattern of cell growth was observed and determined using crystal violet staining. The effects of Y-27632 on the proliferative ability of bovine corneal endothelial cells (BCECs) were assessed according to 5-bromo-2'-deoxyuridine (BrdU) cell incorporation. After BrdU was incubated for an additional 24 h, the proliferation was assessed based on BrdU incorporation. Photographs of multiwell plates stained with crystal violet exhibited a cellular growth pattern similar to that of the TrypLE-preparations. The number of colonies increased from day 1 to day 7 in the BCECs with or without Y-27632, prepared with either TrypLE or collagenase A (A, C). The BrdU incorporation rates increased from day 1 to day 3 and decreased after day 4. There was no statistical difference between the groups prepared with TrypLE from day 1 to day 6. However, Y-27632 significantly decreased the BrdU incorporation rates on day 7 and day 8 (B). In contrast, the BrdU incorporation rates remained essentially constant in the BCECs with or without Y-27632 prepared with collagenase A from day 1 to day 7, and differed from those in the TrypLE-prepared BCECs (B,D). * indicates p<0.05.