Mammalian cell fusion in an electroporation device.
We and others have been interested in the phenomenon of gene 'extinction' in somatic cell hybrids, reasoning that the study of this process is likely to reveal underlying mechanisms responsible for limiting the expression of specialized genes only to appropriate cell types. In the course of our studies in this area, we have developed a simple and economical method of fusing mammalian cells, using an electroporation device. In fusions between murine myeloma and T lymphoma lines, hybrid cell recoveries were typically one per 10(5) [corrected] input myeloma cells. Because of our interest in the regulation of immunoglobulin heavy chain (IgH) gene expression, we analyzed the hybrids for both IgH gene composition and expression. The hybrid lines were phenotypically indistinguishable from those generated by the more conventional, polyethylene glycol (PEG)-induced fusion protocol. There was a notable increase, however, in the number of hybrids that retained IgH-encoding chromosomes from both parental lines.
Radomska
H S
HS
Department of Biological Sciences, Hunter College, City University of New York, NY 10021, USA.
Eckhardt
L A
LA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D002459:Cell Fusion; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D018274:Electroporation / Q000379:methods*; D005803:Genes, Immunoglobulin; D006822:Hybrid Cells; D006825:Hybridomas / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000032:analysis / Q000235:genetics; D051379:Mice; D008806:Mice, Inbred AKR; D008807:Mice, Inbred BALB C; D051381:Rats; D013601:T-Lymphocytes / Q000276:immunology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D007143:Immunoglobulin Heavy Chains