Quantifying the syncytialisation of human placental trophoblast BeWo cells grown in vitro.
We have generated lines of BeWo cells that constitutively and stably express either histone H2B tagged with the green fluorescent protein (GFP), or the mitochondrial targeting sequence of subunit VIII of cytochrome c oxidase fused with a red fluorescent protein; one line has nuclei that fluoresce green, the other mitochondria that fluoresce red. Expression of these tagged proteins has no effect on the rates of DNA, RNA and protein synthesis, or on the amounts of human chorionic gonadotropin (hCG) secreted after treatment with forskolin. We used fluorescence-activated cell sorting (FACS) to monitor the extent of cell fusion (syncytialisation) between these two lines; fused cells are readily and accurately detected by their green/red fluorescence. This assay should prove useful in the investigation of the molecular mechanisms involved in trophoblast syncytialisation.
Kudo
Yoshiki
Y
Department of Human Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK. yoshiki.kudo@anat.ox.ac.uk
Boyd
C A R
CA
Kimura
Hiroshi
H
Cook
P R
PR
Redman
C W G
CW
Sargent
I L
IL
D002459:Cell Fusion / Q000379:methods*; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins; D006657:Histones / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007930:Leucine; D008164:Luminescent Proteins / Q000235:genetics; D024101:Mitochondrial Proteins / Q000096:biosynthesis / Q000235:genetics; D013936:Thymidine; D014162:Transfection; D014316:Tritium; D014327:Trophoblasts / Q000166:cytology / Q000502:physiology*; D014529:Uridine; D000097573:Red Fluorescent Protein
D005576:Colforsin; D049452:Green Fluorescent Proteins; D006657:Histones; D007930:Leucine; D008164:Luminescent Proteins; D024101:Mitochondrial Proteins; D013936:Thymidine; D014316:Tritium; D014529:Uridine; D000097573:Red Fluorescent Protein