Maintaining placental syncytiotrophoblast, a specialized multinucleated transport epithelium, is essential for normal human pregnancy. Syncytiotrophoblast continuously renews through differentiation and fusion of cytotrophoblast cells, under paracrine control by syncytiotrophoblast production of human chorionic gonadotropin (hCG). We hypothesized that K⁺ channels participate in trophoblast syncytialisation and hCG secretion in vitro. Two models of normal term placenta were used: a) isolated cytotrophoblast cells and b) villous tissue in explant culture. Cells and explants were treated with K⁺ channel modulators from 18h, and day 3, onwards respectively. Culture medium was analyzed for hCG, to assess secretion, and lactate dehydrogenase (LDH), to indicate cell/tissue integrity. hCG was also measured in cytotrophoblast cell lysates, indicating cellular production. Syncytialisation of cytotrophoblast cells was assessed by immunofluorescent staining of desmosomes and nuclei. Over 18-66h, mononucleate cells fused to form multinucleated syncytia, accompanied by a 28-fold rise in hCG secretion. 1mM Ba²⁺ stimulated cytotrophoblast cell hCG secretion at 66h compared to control, whereas 5mM tetraethylammonium (TEA) inhibited hCG secretion by >90%. 0.1-1mM 4-aminopyridine (4-AP) reduced cytotrophoblast cell hCG secretion, and elevated cellular hCG; without altering cellular integrity or syncytialisation. In villous explants, hCG secretion was not altered by 1mM Ba²⁺, but inhibited by 5mM 4-AP and 5/10mM TEA, without affecting LDH release. Anandamide, pinacidil and cromakalim were without effect in either model. In conclusion, 4-AP- and TEA-sensitive K⁺ channels (e.g. voltage-gated and Ca²⁺-activated) regulate trophoblast hCG secretion in culture. If these K⁺ channels participate in hCG secretion in situ, they may regulate trophoblast turnover in health and disease.