HYPOXIA-REOXYGENATION MODIFIES THE LIPID PROFILE AND DISRUPTS SYNCYTIALIZATION IN BEWO CELL LINE

ETCHEVERRY TOMAS; MATE SABINA; MARTÍNEZ NORA; VAILLANCOURT CATHY ; FARINA MARIANA

Simposio; 9th Latin American Symposium on Maternal-Fetal Interaction and Placenta (IX SLIMP); 2022

Objectives: To study the effect of hypoxia-reoxygenation (HR) on syncytialization of BeWo cell line and evaluate its impact on the lipid composition of the membrane, focusing on poly-unsaturated fatty acids (PUFAs).Methods: Cell culture: BeWo cell line was cultured for 72h in DMEM/F-12 + 10% FBS at 37°C in a 5% CO2, 95% air atmosphere in the absence or presence of 25μM Forskolin (FK) to induce syncytialization. For the hypoxia-reoxygenation group, cells were exposed to three hypoxic events of 4h each: at 0h, 24h, and 48h. Hypoxia was generated by incubation in a hypoxia chamber with 2% O2, 5% CO2, 93% N3. Culture media were changed daily. Human chorionic gonadotropin (hCG) was quantified in the conditioned media by ELISA. GCM1 mRNA levels were measured by RT-qPCR.Lipidic quantification and profiling were carried out by GLC. Caveolin-1 protein levels were evaluated by WB.Results: Hypoxia-reoxygenation significantly diminishes hCG secretion in forskolin-stimulated cells, suggesting an impairment in syncytialization (n=5, p