title: A non-invasive technique for quantifying and isolating fused cells.
abstract: Cell-cell fusion is an important biological and pathological event. There are limited techniques for studying both the process of cell-cell fusion and the fate of fused cells. We have developed a non-invasive assay for the temporal analysis of cell-cell fusion, quantification of fused cells, and isolation of fused cells. Briefly, cells are transfected with either the T7 bacteriophage RNA polymerase, or yellow fluorescent protein (YFP) driven by a T7 specific promoter. Cells are mixed and induced to fuse. When cells expressing T7 RNA polymerase and T7 promoter driven YFP (T7-YFP) fuse and the cellular contents mix, the YFP is expressed. These YFP-positive cells can be detected with a fluorescent microscope, quantified by flow cytometry, or collected using fluorescence associated cell sorting. Isolated YFP-positive cells can be monitored to determine the fate of fused cells, specifically for the rates of growth, transformation, and changes in chromosome number.
authors.affiliation: Department of Cell Biology, University of Oklahoma Health Sciences Center, College of Medicine, 940 Stanton L. Young Blvd, Biomedical Sciences Building, Rm 553, Oklahoma City, OK, 73104, USA.
authors.forename: Lulin
authors.initials: L
authors.lastname: Hu
authors.forename: Kendra
authors.initials: K
authors.lastname: Plafker
authors.forename: James
authors.initials: J
authors.lastname: Henthorn
authors.forename: Brian P
authors.initials: BP
authors.lastname: Ceresa