A two-colour fluorescence assay for the measurement of syncytial fusion between trophoblast-derived cell lines.
Syncytial fusion is a key event in implantation and placentation. Its regulation is only poorly understood. We present a cell-cell fusion assay based on staining of cells in two portions with a green and a red fluorescent cytoplasmic dye that become intracellularly mixed only after syncytial fusion. We quantified cell-cell fusion by fluorescence microscopy in choriocarcinoma cell lines BeWo, JAR and JEG3 and in some non-trophoblastic cell lines and found clear differences in fusion behaviour. Only BeWo cells fused with each other, while the other cell lines tested did not. BeWo cells also fused with all other cell lines tested. The efficiency of cell-cell fusion of BeWo cells was stimulated by forskolin. We tried to correlate messenger levels of syncytin and its receptor RDR with the fusion index of choriocarcinoma cells. BeWo and JAR cells contained readily detectable and forskolin-inducible levels of syncytin mRNA, whereas this messenger was barely detectable in JEG3 cells. RDR transcript levels were similar in all cell lines tested and were unaffected by forskolin treatment. The data suggests that the expression of syncytin and RDR messengers alone does not guarantee successful fusion. The fusion assay presented in this paper is a useful tool to study syncytial fusion in an accurate and quantitative way.
Borges
M
M
Department of Anatomy II, University Hospital Aachen, Wendlingweg 2, D-52057 Aachen, Germany.
Bose
P
P
Frank
H-G
HG
Kaufmann
P
P
Pötgens
A J G
AJ
D002459:Cell Fusion; D002460:Cell Line; D045744:Cell Line, Tumor; D003116:Color; D005456:Fluorescent Dyes; D015726:Giant Cells / Q000166:cytology*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D012313:RNA / Q000378:metabolism; D014327:Trophoblasts / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure
D005456:Fluorescent Dyes; D012313:RNA