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To establish a simple and quantitative live cell fusion assay for placental syncytialization, we generated stable GFP and dsRed expressing fusogenic BeWo cell lines. Fluorescent Activated Cell Sorting was shown to provide a quantitative determination of forskolin (cAMP-mediated) fusion in a time and concentration dependent manner consistent with the increased secretion of beta human chorionic gonadotrophin (β-HCG) and appearance of multi-nucleated cells. Analyses of the fusion process demonstrated that in addition to increased cAMP levels, simultaneous reduction of intracellular calcium and inhibition of Type 1 phosphatidylinositol 3 kinase (PI3K)/Akt signaling also resulted in cell fusion. Although individual blockade of calcium channel function or PI3K/Akt signaling was without effect, the combination with forskolin resulted in a potentiation of cell fusion. These data demonstrate syncytialization is a complex process that depends upon the regulation of distinct signaling inputs that function in concert with each other.

Original publication




Journal article


PLoS One

Publication Date





Blotting, Western, Calcium, Calcium Channels, Cell Fusion, Cell Line, Tumor, Choriocarcinoma, Chorionic Gonadotropin, beta Subunit, Human, Class I Phosphatidylinositol 3-Kinases, Colforsin, Cyclic AMP, Female, Flow Cytometry, Humans, Lentivirus, Microscopy, Confocal, Placenta, Polymerase Chain Reaction, Pregnancy, Proto-Oncogene Proteins c-akt