TINGKAT PERKEMBANGAN AWAL EMBRIO SAPI IN VITRO MENGGUNAKAN MEDIA TUNGGAL BERBAHAN DASAR TISSUE CULTURE MEDIUM (TCM) 199 Early Bovine Embryonic Development Rate in Vitro Using Single Medium Based on Tissue Culture Medium (TCM) 199

Research on early bovine embryonic development in vitro has been done. Oocytes were collected by aspiration technique from slaughter house ovarian and classified based on the number of cumulus cell layers and homogenous of cytoplasm. Maturation media consisted of tissue culture medium (TCM) 199 supplemented with 10 IU/ml pregnant mare’s serum gonadotropin (PMSG), 10 IU/ml human chorionic gonadotropin (hCG), and 10% fetal bovine serum (FBS). Maturation were done in 5% CO2 incubator 39 C for 24 h. Fertilization was done using two different media namely routine fertilization media and media based on TCM 199 supplemented with BSA and heparin. After fertilization periode, oocytes were denuded and then cultured in TCM 199 supplemented with essential and nonessential amino acid and 10% FBS for three days. Maturation, fertilization, and culture were done in 5% CO2 incubator 39 C. Results of the experment revealed that number of mature oocytes (MII) in our system reaches 81.5%. Meanwhile number of embryonic development were higher in routine media compared to media based on TCM 199 (44.4% vs 23.2%). However, there were no signifcant difference in the number 4-8 cells embryo in both media. It is concluded that single media based on TCM 199 could be used for in vitro embryo production. However further research are required on addition comppound to improve fertilization and embryonic development in the media based on TCM 199. ____________________________________________________________________________________________________________________