Analyzing Trophoblast Fusion Using Immunofluorescence and Split Protein Complementation Assays

The fusion of cytotrophoblasts into a multinucleated syncytiotrophoblast is essential for placental development. For studies investigating syncytiotrophoblast formation, various methods are available to analyze the fusion efficiency of trophoblast cells in vitro. Here, we describe protocols for measuring trophoblast fusion using immunofluorescence and an assay employing complementary parts of a split green fluorescent protein that self-reassociates and generates a fluorescent signal following cell fusion. Together, these approaches allow for a comprehensive and robust analysis of the fusion index in trophoblast cells and can strengthen the accuracy and throughput of investigations into factors that may regulate syncytiotrophoblast development.