Intramuscular injection of increasing doses of Methotrexate Folateantagoniste (25, 50, 100, 150, 250, 300 mg/ kg) on Swiss strain males mice , resulted in decreasing Spermatogenesis at stage VII whereas higher doses had lethal effects. In fact doses of 25 and 50 mg per kg were tolerable by mice and the cytotoxic tolerance level decreased at higher doses (100 and 150 mg/kg). Acute cytotoxic effect was noted at doses of 200 mg/kg and resulted in destruction of cell line. Doses of 250 and 300 mg/kg had lethal effect and the mice died 1 week after injection. In conclusion Methotrexate had cytotoxic effects on sperm cell lines. 100mg/kg doses decreased spermatogenesis while 200 mg/kg doses caused greater side effects and higher doses were lethal to the mice 1 week after injection.