Methotrexate is one of the most important and practical drugs for the treatment of cancer. One of the important side effects of this drug is the development of various infertility in males. This study aimed to evaluate the effect of medium conditioning of cultured Sertoli cells on methotrexate-induced damage in the testicular tissue of mice. In this experimental study, 24 adult male mice were randomly divided into 3 groups of 8. The first group received physiological serum intraperitoneally during the experiment. The second group received methotrexate at a dose of 10 mg/kg once a week for two weeks and the third group received methotrexate at a dose of 10 mg/kg once a week for two weeks. Along with the conditioned culture medium, Sertoli cells were received 40 days after the last methotrexate injection. Fifty days later, the animals were anesthetized intraperitoneally by injection of ketamine and xylazine. After blood sampling, testicular tissue was obtained and placed in 10% formalin. The results of this study showed that methotrexate has destructive effects on testicular tissue and causes seminiferous tubular atrophy, decreased epithelial thickness, increased interstitial tissue thickness, and decreased levels of sex hormones including FSH, LH, and testosterone compared to the control group. Based on this study, it can be concluded that the use of medium conditioning of Sertoli cells reduces the damage caused by methotrexate on the testicular tissue and the Sex hormone levels (FSH, LH, and testosterone) bring male mice closer to the physiological state.
