US 37987295 A description dulbecco 39 s minimal eagle 39 s medium fetal calf serum glutamine penicillin and streptomycin were purchased from quality biological inc gaithersburg md hanks 39 balanced salt solution and trypsin edta were obtained from gibco laboratories grand island n y 1 6 diphenylhexatriene and tetrahydrofuran were from aldrich chemical co inc milwaukee wis gossypol and gossypol acetic acid were gifts from the national research institute for family planning beijing china the established line of small cell human adrenocortical carcinoma sw 13 was purchased from the american type culture collection rockville md sw 13 cells were seeded in a 25 cm 2 tissue culture flask costar cambridge mass at densities of 1 u00d7 10 4 cells 5 ml of dulbecco 39 s minimal eagle 39 s medium supplemented with 10 fetal calf serum 100 ug ml streptomycin 100 units ml penicillin and 2 mm glutamine the cells were grown in a humidified 37 u00b0 c incubator with a 5 co 2 95 air atmosphere a gossypol stock solution in absolute ethanol was added to the culture medium to yield final concentrations of 0 0 5 5 and 50 um gossypol with a 0 1 final ethanol concentration after 1 2 4 or 6 days of incubation the culture medium containing a few floating cells was removed adherent cells were trypsinized.