The purpose of this paper is to study the method of collagenase-and BSA desity gradients, wehave established a serum-free culture for rhesus monkey Sertoli cells and studied the estradiolregulation of hFSH oFSH, db-cAMP, MIX, Cholera tixon and Forskolin in cultured rhesusmonkey Sertoli cells. The results that concentration of hFSH and oFSH were found to be ineffective, but the same condition, db-cAMP, MIX, Cholera tixon and Forskolin stimulated the conversion of androstenedione to estradiol by Sertoli cells of rhesus monkey.