CONICET | Buscador de Institutos y Recursos Humanos

Objective:The mitochondrial bioenergetics constitute an important topic in the development of placenta dueto the metabolic demand throughout gestation and may determine the correct trophoblastdifferentiation, from cytotrophoblast (CT) to syncytiotrophoblast (SCT). In this sense, we test thefollowing hypothesis, if mitochondrial metabolism is different for the two cell subpopulations oftrophoblast, in a cell culture model in vivo of syncytialization of placental villous trophoblast, themembrane potential will be changed in the cells after differentiation. Our aim was to evaluate themembrane potential BeWo cells differentiated and undifferentiated.Methods:To evaluate the effect of differentiation on the membrane potential in BeWo cells induces by 25μMforskolin (FSK), fluorescence microscopy analysis of mitochondrial membrane potential after dualloading with TMRE and Hoechst was performed. For the assay, 1x106cells per well were cultured ina 6-well plate until they reached semi-confluence. The cell-permeant MitoTracker® probes containa mildly thiol-reactive chloromethyl moiety was used for labeling of mitochondria, TMRE (Excited at549 nm, emits at 574 nm) RED, use concentration 1 μM; and the total labeling of nuclei was carriedout 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 wereobserved through fluorescence microscopy, indicating that both subpopulations are metabolicallydifferent.Conclusions:Our results may suggest that these differences in the mitochondrial metabolism between theundifferentiated and differentiated cell it could indicate very important participation of thisorganelle in the trophoblast cells differentiation.

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