Abstract
Placental oxidative stress is present throughout the duration of pregnancy, but it is when oxidative stress exceeds the normal physiological level that complications can occur. Trophoblast cell lines are commonly utilized for oxidative stress research due to their distinct uniform cell population and easy‑to‑apply interventions. However, conflicting results are often reported when different oxidative stress cell models are used. In this study, the aim was to characterize the intracellular and extracellular metabolite profiles of different oxidative stress cell models commonly used in the research of pregnancy complications. HTR8/SVneo human trophoblast cell lines were treated with five different oxidative stress‑inducing conditions: Hypoxia (1% oxygen); hypoxia and reoxygenation; cobalt chloride (CoCl2; 300 µmol/l); sodium nitroprusside (SNP; 2.5 mmol/l); and the serum of women with preeclampsia (10% v/v). Intracellular metabolites were extracted from cells and extracellular metabolites were collected from spent media for metabolomic analysis via gas chromatography‑mass spectrometry. The results demonstrated that there were distinct differences in the intracellular and extracellular metabolome between the different cell models. Meanwhile, treatments with exogenous drugs, such as CoCl2 and SNP, resulted in more similar metabolite profiles. These disparities between the different oxidative stress cell models will have implications for the applications of these results, and highlight the need for the standardization of oxidative stress cell models in obstetric research.