Placental hypoxia has been implicated in pregnancy pathologies, including fetal growth restriction and preeclampsia, however the mechanism by which the trophoblast cell responds to hypoxia has not been adequately explored. Glucose transport, a process crucial to fetoplacental growth, is upregulated by hypoxia in a number of cell types. We investigated the effects of hypoxia on the regulation of trophoblast glucose transporter expression and activity in BeWo choriocarcinoma cells, a trophoblast cell model, and human placental villous tissue explants. GLUT1 expression in BeWo cells was upregulated by the hypoxia-inducing chemical agents, desferroxamine and cobalt chloride. Reductions in oxygen tension resulted in dose-dependent increases in GLUT1 and GLUT3 expression. Exposure of cells to hypoxic conditions also resulted in an increase in transepithelial glucose transport. A role for HIF-1 was suggested by the increase in HIF-1 as a result of hypoxia and by the increase in GLUT1 expression following treatment of BeWo with MG-132, a proteasomal inhibitor which increases HIF-1 levels. The function of HIF-1 was confirmed in experiments where the hypoxic upregulation of GLUT1 and GLUT3 was inhibited by antisense HIF-1. By contrast with BeWo, hypoxia produced minimal increases in GLUT1 expression in explants, however treatment with MG-132 did upregulate syncytial basal membrane GLUT1. Our results show that glucose transporters are upregulated by hypoxia via a HIF-1-mediated pathway in trophoblast cells and suggest that the glucose transporter response to hypoxia in vivo will be determined not only by low oxygen tension but also by other factors which modulate HIF-1 levels.