In the present study, we examined sperm quality derived from epididymis in Hanwoo bull at 13 months (before puberty). Collected semen from epididymis of two bulls were cryopreserved in LN2. After thawing of frozen semen was examined sperm motility and sperm motility parameters by Computer Assessment of Sperm Analysis (CASA) system. Progressive motility of frozen-thawed sperm in epididymis was lower than that of before freezing. Curvilinear velocity, straight-line velocity, average path velocity and linearity of frozen-thawed sperm in epididymis were lower than those of before freezing. In addition, blastocyst development of oocytes fertilized with frozen-thawed sperm from epididymis of bulls were examined. Blastocyst development rates after fertilization in vitro with frozen-thawed sperm between epididymis and commercial semen were similar (38.5 vs. 32.0, respectively). In conclusion, sperm derived from epididymis in pre-puberty of Hanwoo bulls have low motility and many are not yet mature; however, it has fertilizing ability and blastocyst development after fertilization.