The present study was undertaken to determine the ability of cultured luteal cells from human corpora lutea to secrete progesterone (P<sub>4</sub>) and prostaglandins (PGs), and to assess the effects of the products of the lipoxygenase pathway on luteal P<sub>4</sub> production. Luteal cells responded to human chorionic gonadotropin (hCG) with a significant increase (2- to 7-fold) in P<sub>4</sub> production. Arachidonic acid significantly stimulated PGE<sub>2</sub> synthesis by luteal cells in a dose-dependent manner. Both basal PGE<sub>2</sub> production and the responsiveness to arachidonic acid were maintained for 8 days. In contrast, both PGF<sub>2α</sub> and 6-keto-PGF<sub>1α</sub> production abruptly declined as the culture proceeded. However, the addition of hCG did not further stimulate the accumulation of the 3 PGs assayed. In the subsequent experiment, 5-hydroxyeicosatetraenoic acid (5-HETE) and the reaction products of soybean lipoxidase of arachidonic acid (AA-LIP) were utilized for evaluating the involvement of the lipoxygenase pathway in luteolysis. The addition of 5-HETE dose-dependently inhibited P<sub>4</sub> production by the cultured luteal cells. Although treatment with either arachidonic acid or lipoxidase alone had no effect on P<sub>4</sub> production, AA-LIP significantly reduced P<sub>4</sub> production in the presence or absence of hCG. These results suggest that the products of the lipoxygenase as well as of the cyclo-oxygenase pathway may be important in regulating the life span and function of human corpora lutea.