Gonadotrophins and various growth factors, but not sex steroids, can induce resumption of meiosis in vitro, but only in oocytes enclosed by cumulus-granulosa cells. Follicular purines prevent resumption of meiosis. This process can be overcome, in vitro, by a transient elevation of cyclic AMP resulting in the production of a diffusible meiosis-inducing substance secreted by the cumulus cells. A meiosis-inducing activity has been detected in gonads of different species, for example, in preovulatory follicular fluid of women and in mouse testes. We report here the isolation and characterization of meiosis-activating sterols from human follicular fluid and bull testes and the synthesis of two closely related C29 sterols. All these sterols induce a resumption of meiosis in cultured cumulus-enclosed and naked mouse oocytes indicating their nonspecificity across species and sex. This family of sterols is for the first time considered crucial to meiosis.