Recombinant human-derived hyaluronidase has been reported to have the advantage of no risk of viral infection unlike bovine-derived hyaluronidase. Bovine-derived hyaluronidase and recombinant human-derived hyaluronidase were used for oocyte denudation and the results were compared. ICSI patients from whom four or more oocytes were collected between August 2008 and December 2008 were investigated, and a total of 376 oocytes were collected from 39 subjects. The oocytes were divided into groups based on treatment: bovine hyaluronidase (group A), and recombinant hyaluronidase (group B). The time required for oocyte denudation was slightly longer in group B than in group A, but there were no significant differences between groups A and B in the fertilization rate (70.9% vs 68.0%), the denaturation rate (9.5% vs 7.8%), the division rate (92.4% vs 97.1%), the rate of good quality embryos per embryo development (63.9% vs 71.3%), the rate of blastocyst formation (41.7% vs 45.7%), or the rate of good quality blastocysts per blastocyst formation on day 5 (25.0% vs 44.8%).