Formation of C19 11-hydroxysteroids by porcine Leydig cells

In a previous study, we reported the presence of 11 beta-hydroxyandrostenedione and 11 beta-hydroxytestosterone in testicular vein blood from mature male pigs. Since C19 steroids with an oxygen function at C11 have not been recorded as products of steroid biosynthesis in normal mammalian testes, we have examined their possible production in purified preparations of porcine Leydig cells. Both androstenedione and cortisol were added as substrates in studies using cell incubations of Leydig cells from mature boars (greater than 8 months old). Steroids were recovered from media by solid-phase extraction and separated by reversed-phase high performance liquid chromatography. Peaks corresponding to retention times of authentic standard steroids were seen for both 11 beta-hydroxyandrostenedione and 11 beta-hydroxytestosterone from each substrate. Generally, lesser amounts of C19 11-oxosteroids were noted also. Definitive confirmation was made by gas chromatography - mass spectrometry for 11 beta-hydroxyandrostenedione in the media.