Hyperosmolarity induced a biphasic arrest in corneal epithelial cells. hTCEpi cells were cultured in basal media with increasing concentrations of salt. Cell number and cell cycle were assessed at six and 24 hours. (A) There were no differences in cell number between any of the test conditions at six hours (P = 0.093, one-way ANOVA, n = 3). (B) After 24 hours, there was a progressive decrease in cell number with increasing osmolarity (*P < 0.001, one-way ANOVA, SNK multiple comparison test, n = 3). (C) After six hours in hyperosmolar culture, hTCEpi cells were arrested at G2/M (*P < 0.001, **P = 0.044, one-way ANOVA, SNK multiple comparison test, n = 3). (D) At 24 hours, hTCEpi cells cultured under hyperosmolar conditions were arrested in G0/G1 (*P < 0.001, ***P = 0.004, ****P = 0.002, one-way ANOVA, SNK multiple comparison test, n = 3). Data represented as mean ± standard deviation. Graphs representative of a single experiment repeated 3 times. KBM, keratinocyte basal media. Number indicates mOsM.