Quantification of cell fusion by flow cytometry.
Cells of different types can be induced to fuse by electroshock. Cells of one type are typically dominant and are able to reprogram the nuclei derived from cells of the other type, in fusion hybrids derived from one cell of each type. Flow cytometry provides a quick and objective technique to assess cell fusion for nuclear reprogramming studies. Two cell types are each stained with a different fluorescent dye and then induced to fuse to form fusion products called heterokaryons. Heterokaryons can be identified and quantified by flow cytometry as double-stained events. Protocols are provided for the optimization of cell staining under conditions that minimize cell clumping and dye leakage. If spectral overlap occurs between emission spectra of the two stained cell types, the data will need to be electronically compensated.
Sullivan
Stephen
S
Wellcome Trust/Cancer Research United Kingdom Gurdon Institute of Cancer and Developmental Biology, University of Cambridge, United Kingdom.
Waterfall
Martin
M
Gallagher
Ed J
EJ
McWhir
Jim
J
Pells
Steve
S
D000818:Animals; D002459:Cell Fusion; D002467:Cell Nucleus / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000166:cytology; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006822:Hybrid Cells / Q000166:cytology*; D051379:Mice; D008808:Mice, Inbred CBA; D008954:Models, Biological; D013234:Stem Cells / Q000166:cytology; D013950:Thymus Gland / Q000378:metabolism