Mammalian cell fusion in an electroporation device

• Published in: Journal of immunological methods Vol. 188; no. 2; pp. 209 - 217
• Main Authors: Radomska, Hanna S., Eckhardt, Laurel A.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 27-12-1995; Elsevier
• Subjects: Animals; Antibodies, Monoclonal - biosynthesis; Biological and medical sciences; Cell Fusion; Cell Line; Cell Survival; Cells, Cultured; Diverse techniques; Electroporation - methods; Fundamental and applied biological sciences. Psychology; Genes, Immunoglobulin; Hybrid Cells; Hybridoma; Hybridomas - immunology; Immunoglobulin Heavy Chains - analysis; Immunoglobulin Heavy Chains - genetics; Mice; Mice, Inbred AKR; Mice, Inbred BALB C; Molecular and cellular biology; Myeloma; Rats; T lymphoma; T-Lymphocytes - immunology; Tumor Cells, Cultured; Hybridoma; T lymphoma; Cell fusion; Myeloma; Vertebrata; Cell line; Mammalia; Mouse; T-Lymphocyte; Rodentia; Electroporation; Method; Lymphoma
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/0022-1759(95)00205-7

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 105 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.