A non-invasive technique for quantifying and isolating fused cells.
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.
Hu
Lulin
L
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.
Plafker
Kendra
K
Henthorn
James
J
Ceresa
Brian P
BP