In vitro Embryo Production and Transfer in Cattle

TCM-199 + 10% Fetal Calf Serum (FCS) + 2μg/ml FSH and 20-22 hours incubation period were used for in vitro maturation. Direct washing method by Brackett and Oliphant (BO) medium and 5 or 6 hours incubation period were used for in vitro fertilization. Charles Rosencrans (CR1aa) medium was used for in vitro emryo culture. Oocytes incubation was done in 39 ºC, 5 % CO₂ and over than % 95 humidity for each IVM, IVF and IVC procedures. 230 ovaries were collected after 12 visited to sloughterhouse. 1661 A and B quality oocytes and 498 C quality and degenerated oocytes were collected by aspirating and slicing of ovaries. 7.22 A and B quality oocytes and 2.16 C quality and degenerated oocytes totally 9.38 oocytes provided from per ovaries. Only A and B quality oocytes were used for production of in vitro embryos. Maturation rate was 92.0 %, cleavage rate was 72.0 % and coming into morulae-blastocyst stage rate was 34.0 %. Killed spermatozoa were used to diagnosis of the parthenogenetical cleavage rate. Parthenogenetic cleavage rate was 4.5 % after taken some oocytes sample randomly. In vitro produced embryos were transferred to five recipient cows. Two pregnancies were diagnosed by ultrasound imaging in the 50th day after transfers then one healthy calf delivered. Keywords : In vitro, Embryo, Maturation, Fertilization, Cattle, Transfer