Figure 4
The use of Y-27632 for cell survival during regular cellular passage and recovery from cryo-preservation.
(A) Confluent cultures of CECs maintained in M5-Endo medium, with or without a 1-hour pre-treatment of 10 μM Y-27632 were analyzed for overall percentage of live cells, as gauged by the Annexin V detection kit, following regular cellular passage - ‘C’ denote control; ‘Y’ denote Y-27632 treated (n = 6). (B) Overall percentage live cells were also assessed for both groups of cryo-preserved CECs that were either untreated or those that were exposed to 10 μM Y-27632 - ‘C’ denote control; ‘Y’ denote Y-27632 treated (n = 6). Representative micrographs of (C) cryo-preserved control or untreated CECs and (D) corresponding CECs that were exposed to 10 μM Y-27632 that were thawed out and seeded at physiological density of approximately 3,000 cells per mm2. (C′) and (D′) depicts CECs from corresponding micrographs (C) and (D) that were briefly treated with TrypLE Express for 5 seconds to better visualize the CECs. Scale Bars: 50 μm.
