By using aspiration from the vas deferens, apparently good quality spermatozoa can be obtained for in-vitro fertilization (IVF) in cases of non-treatable anejaculation. Being squeezed from the epididymis during aspiration, the spermatozoa may be immature and their fertilizing capacity lower than that of ejaculated spermatozoa. Our case report describes a couple who achieved pregnancy when intracytoplasmic sperm injection (ICSI) was carried out with frozen-thawed spermatozoa aspirated from the vas deferens of a man whose anejaculation was associated with diabetes mellitus. In the aspiration, 50 x 10(6) spermatozoa were obtained. One half of them was frozen, and the other half was used fresh for conventional IVF, resulting in total fertilization failure of all the oocytes. The second treatment was ICSI, in which eight out of 11 oocytes injected with frozen-thawed spermatozoa showed normal fertilization. The second frozen embryo transfer resulted in a normal pregnancy.