The aim of this study was to evaluate the timed AI (TAI) pregnancy rate using fresh sexed semen (15 to 20ºC). The work was carried out in MS state, where 567 zebu females (269 heifers and 298 cows similarly distributed among the groups) were synchronized for TAI. On D0, 2 mg of EB (Gonadiol®; Zoetis, São Paulo, SP, Brazil) was injected and an intravaginal P4 device (DIB ®, 1g of P4; Zoetis) was inserted. At D9, the P4 device was removed and 0.6 mg of EC (ECP®, Zoetis, Brasil), 12.5 mg of Dinoprost (Lutalyse®; Zoetis) and 300 IU of eCG (Novormon®; Zoetis) on cows and 200IU of eCG on heifers was injected. On day 11, 48 hours (149 cows and 134 heifers) or 54 hours (149 cows and 135 heifers) after removal of the P4 device, the females were inseminated, using 6 Nellore bulls (similarly distributed among the groups) collected on the same day of P4 device removal (part of the ejaculate was frozen conventional and part sexed using flow cytometry technology for later cooling). At the time of AI (between 24 and 36 hours after leaving the semen from the laboratory), the animals were divided into four treatments in a 2x2 factorial design: conventional frozen semen 48h (CF48; N=143), conventional frozen 54h (CF54; N=128), fresh sexed semen 48h (FS48; N=140) and fresh sexed semen 54h (FS54; N=156). Storage during transport took place in a container developed for this purpose, with controlled temperature throughout the laboratory output and the TAI (15 to 20ºC). The pregnancy rate was evaluated 30 days after AI, and gender proportion 80 days after TAI, by transrectal ultrasound. The data were analyzed using SAS PROC GLIMMIX (SAS/STAT® 9.2). There was no interaction bull*treatment for P/AI (P=0.2689). Also, no interaction treatment*moment of TAI was found for P/AI (P= 0.2777). The P/AI according to the groups was: CF48: 57.3%; CF54: 66.4%; FS48: 52.1%; FS54: 56.4%. There was no effect of the type of semen used in P/AI (Conventional: 61.6%; Fresh sexed: 54.4%; P= 0.0956). Furthermore, the TAI 48h or 54h after P4 device removal presented same P/AI (48h: 54.8%; 54h: 60.9%; P= 0.1130). However, there was difference between heifers; 52.8% and cows; 62.4% (P=0.0282). There was a difference in the proportion of pregnancies of the desired gender (females) according to the treatment (Conventional: 53.5%; Fresh sexed: 94.1%; P<0.001). However, there was no effect on the proportion of the desired gender according to AI time (P= 0.0809), nor in relation to cows x heifers (P=0.8072), and the bull used (P=0.6505). The results show that the fresh sexed semen, maintained under the temperature conditions described, reaches P/AI on TAI like conventional frozen semen when used 48h or 54h after P4 device removal (up to 36 hours after release in the laboratory). So, data suggests that it is possible to increase the number of pregnancies of calves of the desired gender without decrease the TAI P/AI and in a near future this technology can be commercially established in the market.