AQP9- mediated lactate transport: an alternative source of energy in placenta.

MEDINA Y.; SZPILBARG N.; REPPETTI J.; DAMIANO A. E.

Simposio; International Symposium on Reproductive Health: overcoming barriers for research in reproduction; 2021

Introduction:The mitochondrial bioenergetics constitute an important topic in the development of placenta due to the metabolic demand throughout gestation. The placenta consists of multiple cell layers, in which the trofoblast differentiates into two cell types, the cytotrophoblast (CT) is formed internally, cells with well-defined limits and very mitotically active. Externally, the syncytiotrophoblast (SCT), a multinucleated tissue that lacks cellular boundaries, differentiates. In the periphery, the differentiating trophoblast forms projections called primary villi made up of both types of cells; the cytotrophoblast is located centrally, while the syncytiotrophoblast is located at the periphery.Thus, for this study, we tested the hypothesis that the mitochondrial metabolism of the two cell subpopulations, CT and SCT, using a placental cell line that has been widely used as an in vitro model for the placenta. The trophoblast-derived choriocarcinoma cell line (BeWo). BeWo cells are regularly used as a cell culture model to mimic in vivo syncytialization of placental villous trophoblast through triggered fusion by forskolin.Objective: To evaluate the membrane potential BeWo cells differentiated and undifferentiated.Materials and method: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. The difference between the two mitochondria metabolism in these differentiation process it could indicate that very important participation of this organelle in the trophoblast cells differentiation.