This experiment was conducted to study the effects of dietary different sesame oil levels on laying performance, egg quality, serum lipid indices, and yolk cholesterol content of laying hens. One hundred and sixty 28-week-old Suqin green-shelled laying hens with a similar laying rate were assigned to 4 groups with 4 replicates in each group and 10 laying hens per replicate. The diet of the control group was no sesame oil, while the diets of experimental groups contained 1%, 2% and 3% sesame oil, respectively. The energy and protein levels in all the diets were the same. The experiment lasted for 42 d. The results showed that compared with the control group, there were no significant differences in laying rate, total egg weight, total feed intake, and feed/egg among all the groups (P>0.05). The average egg weight and egg shape index in the 2% sesame oil group were significantly higher than those in the control group (P<0.05). The Haugh unit in the 3% sesame oil group was significantly higher than that in the control group and 2% sesame oil group (P<0.05). The yolk color in the 2% and 3% sesame oil groups was significantly deeper compared with the control group (P<0.05). The serum cholesterol and triglyceride contents in the 2% sesame oil group were significantly lower than those in the control group and 3% sesame oil group (P<0.05). The yolk cholesterol content in the 3% sesame oil group was significantly higher than that in the other groups (P<0.05). In conclusion, adding 2% sesame oil to the diet can increase average egg weight, egg shape index, yolk color, and yolk weight, while reduce the serum cholesterol and triglyceride contents; without harmful effects are found in laying rate, feed/egg, and other egg quality indices. We suggest that the level of dietary sesame oil is 2%.