This study aimed to investigate the effect of supplementing 300 mg/ kg tea polyphenols (TP) on yolk cholesterol content and production performance of laying hens during the egg-laying period. A total of 600 Roman laying hens aged 24 weeks were randomly divided into two dietary treatment groups. The feeding experiment lasted for 48 weeks. Layers fed basal diet supplemented with 0 (control group) and 300mg/ kg TP (TP group) diet, respectively. The yolk cholesterol content, laying performance, and egg quality were determined at 28, 38, 48, 58, and 68 weeks of age. The yolk cholesterol content in the TP group was significantly decreased at 28-68 weeks of age (p<0.01), compared to the control group. There was a significant increase in laying rate in the TP group at 38 weeks of age (p<0.05), compared to the control group, while no significant differences during the other laying periods were obtained (p>0.05). The FCR significantly decreased in the TP group at 38 weeks of age whereas AEW significantly increased in the TP group at 58 weeks of age (p<0.05). Similarly, the eggshell thickness and eggshell strength in the TP group significantly increased (p<0.05), compared with the control group at 38 weeks of age. The albumen height and Haugh unit significantly increased at 28 weeks of age (p<0.05). In conclusion, the results showed that the diet supplemented with 300 mg/kg TP had positive effects on production performance of layers during the egglaying period, and could lessen yolk cholesterol content significantly at 28-68 weeks of age.