Epidemiological surveys and animal experiments have shown that 2-bromopropane induces oligozoospermia in exposed workers and inhibits spermatogensis in laborratory animals. However, themechanism by which 2-bromopropane exerts its effects is unknown. To this end, we examined the formation of testosterone by the Leydig cells and their survival of these cells in the Presence of differ-ent concentrations of 2-bromopropane in vitro. Leydig cells were isolated following vascular Perfu-sion, enzymatic dissociation and Percoll gradient centrifugation techniques. The cells were cultured in culture dishes. After 8 h, different cultures were exposed to 2-bromopropane at concentrations of 0.01 mmol/L, 0.10 mmol/L and 1.00 mmol/L. In order to stimulate Leydig cells to secrete testos-terone, human chorionic gonadotropin (hCG) was also added. Cell viability was determined using the trypan blue dye exclusion test and cell numbers were counted by hemocytometer. Testosterone secretion was detected by radioimmunoassay. The cell viability decreased after exposure to 2-bromo-propane in a dose-dependent way, but no morphological change was observed. The cell number de-creased in the 2-bromopropane-treated cultures. The secretion of testosterone did not manifest de -tectable changes in the culture treated with 0.10 mmol/L and 0.01 mmol/L of 2-bromopropane;however, it decreased significantly (P < 0. 02) in the Presence of 1.00 mmol/L. Therefore, ourresults strongly suggest that 2-bromopropane may exert its cytotoxic effects on heydig cells in vitro.We speculate that the decrease in the numbers of Leydig cells caused by 2-bromopropane was medi-ated by a feedback mechanism resulting from a lower testosterone concentration. Epidemiological surveys and animal experiments have shown that 2-bromopropane induces oligozoospermia in exposed workers and inhibits spermatogensis in labor laboratory animals. However, themechanism by which 2-bromopropane exerts its effects is unknown. To this end, we examined the formation of testosterone by the Leydig cells and their survival of these cells in the Presence of differ- ent concentrations of 2-bromopropane in vitro. Leydig cells were isolated following vascular Perfu-sion, enzymatic dissociation and Percoll gradient centrifugation techniques. The cells were cultured in culture dishes. After 8 h, different cultures were exposed to 2-bromopropane at concentrations of 0.01 mmol / L, 0.10 mmol / L and 1.00 mmol / L. In order to stimulate Leydig cells to secrete testos-terone, human chorionic gonadotropin (hCG) was also added. Cell viability was determined using the trypan blue dye exclusion test and cell numbers were counted by hemocytometer. Testosterone secretion was detected by ra The cell viability decreased after exposure to 2-bromo-propane in a dose-dependent way, but no morphological change was observed. The cell number de-creased in the 2-bromopropane-treated cultures. The secretion of testosterone did not manifest however, it decreased significantly (P <0.02) in the Presence of 1.00 mmol / L. Therefore, ourresults strongly suggest that 2-bromopropane may exert its cytotoxic effects on heydig cells in vitro. We speculate that the decrease in the numbers of Leydig cells caused by 2-bromopropane was medi- ated by a feedback mechanism resulting from a lower testosterone concentration.