Superfusion in vitro in the study of ovarian steroidogenesis

A method for the continuous superfusion of porcine corpus luteum tissue is described which readily allows both the introduction of regulatory factors to the incubating tissue, and sampling of the tissue. Oestrogen (principally oestradiol) and progestin (principally progesterone) can be measured for up to 24 h in the superfusate from corpora lutea of all ages, and the secretion of both steroids is stimulated by the addition of luteinizing hormone. The pattern of response of both steroids to a pulse of gonadotrophin was similar in that a rapid transient increase in secretion occurred followed some time later by a secondary and more prolonged response. A second pulse of gonadotrophin introduced 6 h after the first also stimulated steroid secretion, indicating that during superfusion in vitro the porcine corpus luteum does not become refractory to the steroidogenic effect of gonadotrophin.