Male infertility is still a problem for in vitro fertilization. The sperm in the damaged testicles are insufficient for fertilization. Ganoderma lucidum (GL) is a natural product as a supportive agent. It is a powerful antioxidant with the polysaccharides it contains. It has been shown nontoxic in rats and mice. In this study, the antioxidant effect of GL on sperm damage was studied in culture. Sperm collected from the epididymis of adult male rats with TESE were incubated with RPMI media which included 10% FBS. Sperm were cultured in high glucose medium at 10, 100, and 1000 μL / ml for GL effect. Sperm motility was assessed at half-hour intervals. When immotility began, it was examined for morphology with quick-diff stainig. It was also fixed for SEM. Oxidative stress with eNOS and apoptosis with TUNEL of sperm were examined. The motile sperm number decreased in the high glucose medium compared to the control medium. The number of abnormal sperm counts doubled. Morphological abnormalities were confirmed by SEM. The number of marked eNOS sperm and the apoptotic number indicated by TUNEL were increased. The GL application returned all parameters in a meaningful way. Sperm maturation in male infertility is an important factor for successful fertilization and live birth. GL at its highest dose inhibited oxidative stress and apoptosis most effectively. GL is commercial product and is used for many reasons. It was thought that the use of GL in patients with infertility problems would be beneficial. [ABSTRACT FROM AUTHOR]