MEMBRANE POTENTIAL ANALYSIS IN UNDIFFERENTIATED AND DIFFERENTIATED BEWO CELLS.

MEDINA, YOLLYSETH; BUSTAMANTE, JUANITA; ETCHEVERRY, TOMÁS; DAMIANO, ALICIA E.; ACOSTA, LUCAS

Amsterdam

Congreso; International Federation of Placenta Associations 2021 (IFPA2021) to be held jointly with the 8th Latin American Symposium on Maternal-Fetal Interaction and Placenta (VIII SLIMP); 2022

International Federation of Placenta Associations (IFPA)

Objective:The mitochondrial bioenergetics constitute an important topic in the development of placenta due to the metabolic demand throughout gestation and may determine the correct trophoblast differentiation, from cytotrophoblast (CT) to syncytiotrophoblast (SCT). In this sense, we test the following hypothesis, if mitochondrial metabolism is different for the two cell subpopulations of trophoblast, in a cell culture model in vivo of syncytialization of placental villous trophoblast, the membrane potential will be changed in the cells after differentiation. Our aim was to evaluate the membrane potential BeWo cells differentiated and undifferentiated.Methods:To evaluate the effect of differentiation on the membrane potential in BeWo cells induces by 25μM forskolin (FSK), fluorescence microscopy analysis of mitochondrial membrane potential after dual loading with TMRE and Hoechst was performed. For the assay, 1x106 cells per well were cultured in a 6-well plate until they reached semi-confluence. The cell-permeant MitoTracker® probes contain a mildly thiol-reactive chloromethyl moiety was used for labeling of mitochondria, TMRE (Excited at 549 nm, emits at 574 nm) RED, use concentration 1 μM; and the total labeling of nuclei was carried out with Hoescht (Excited at 400 nm, emits at 460-490 nm) BLUE. Use concentration 0.5 μM.Results:After BeWo cells differentiation with FSK important changes in mitochondrial polarization were observed through fluorescence microscopy, indicating that both subpopulations are metabolically different.Conclusions:Our results may suggest that these difference in the mitochondrial metabolism between the undifferentiated and differentiated cell it could indicate very important participation of this organelle in the trophoblast cells differentiation.