Sperm treatment method plays a important role in in vitro fertilization (IVF) because it can improve sperm quality and in vitro developmental potential of IVF embryos. Jeju Black Cattle (JBC) is an indigenous species of Korea and IVF system is very important for mass production and industrialization of JBC. In order to find out the most suitable sperm treatment method, the following methods of percoll gradient, triladyl and percoll-triladyl combination (P, T and PT, respectively) were used to sperm treatment. At 2 days after IVF, the fertilization rate was higher in T and in PT groups than in P group. The blastocyst formation rate was significantly increased in T group than P and PT groups. The total cell number was higher in T and PT groups than in P group. The blastocyst formation rate was increased in T group. When we performed IVF after IVM treatment of individual ovaries, in comparison of the development rates of donor and non-donor cows, there was no significant difference. According to grade (1++, 1+, 1, 2), the development rate was the highest in 1++ grade cattle, followed by 1, 1+ and 2 grade. Also, in vitro developmental potential by carcass weight was the highest at the 300 ~ 350 kg, followed by 300 kg or less, 350 ~ 400 kg, then 400 kg more over. As a result, the most effective sperm processing was resulted in PT method, and the development of IVF embryos was more effective when the carcass grade was high and the carcass weight was about 350 kg. This study demonstrates that JBC sperm can be effectively recovered with simple and efficient PT method and from this protocol, high quality embryos can be produced in vitro.