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Objective: To assess the effects of hydrosalpinx fluid on human cytotrophoblast viability and function in vitro. Design: Human cytotrophoblasts obtained from third-trimester placentas were cultured in vitro with hydrosalpinx fluid, and cell viability and protein production were assayed. Setting: A university hospital. Patient(s): Ten hydrosalpinx fluid samples obtained from seven women with clearly diagnosed hydrosalpinges. Intervention(s): Recovery of hydrosalpinx fluid by transvaginal aspiration or at the time of surgery. Main Outcome Measure(s): Cell viability was assessed by the XTT assay. Secretion of trophoblast oncofetal fibronectin (tropho-uteronectin) and β -hCG by cultured trophoblasts was determined by Western blot and ELISA of the culture media. Result(s): With increasing concentrations of hydrosalpinx fluid from 0% to 20%, there was a significant increase in trophoblast cell viability (1.63-fold increase in 20% hydrosalpinx fluid). Likewise, both Western blot and ELISA assays demonstrated a significant increase in trophouteronectin production by trophoblasts with increasing hydrosalpinx fluid concentrations (3.76-fold increase in 20% hydrosalpinx fluid). β -Human chorionic gonadotropin production also increased significantly in the presence of hydrosalpinx fluid (3.31-fold increase in 20% hydrosalpinx fluid). Conclusion(s): These findings suggest that hydrosalpinx fluid improves human trophoblast viability in vitro and enhances the production of tropho-uteronectin and β -hCG by these cells.