BACKGROUND: To improve breeding programs in dairy cattle artificial insemination using sexed sperm has been applied in different farms. OBJECTIVES: To evaluate pregnancy rate and economic benefit of artificial insemination with sexed sperm in Holstein dairy heifers of Shiraz industrial farms. METHODS: The insemination data of 200 heifers in five farms including sperm types (sexed or conventional) and insemination date and times were collected from March 2009 to March 2010. RESULTS: The pregnancy rate of conventional sperms (72.2%) were significantly higher than the sexed sperms in heifers (49.2%; p=0.001). The sexed sperms insemination showed lower fertility in the winter (36.3%) when compared with the autumn values (73.9%; p=0.02). Cost and benefit analysis showed that costs of the first and the second insemination with sexed sperms were higher than the conventional ones. On the other hand, after the second insemination the economic benefit for the sexed sperms was lower than the conventional one. However, the cost and benefit analysis for sexed sperm values showed a trend to the convential ones. CONCLUSIONS: If the fertility rate of the sexed sperm insemination in healthy heifers reach to the 90 percent of the conventional sperm, sexed sperm insemination can be more beneficial than the conventional one after two insemination.