Cephalin as an efficient fusogen in hybridoma technology: can it replace poly ethylene glycol?
In this study we set up a simple, fast, and highly efficient protocol to fuse cells and produce human hybridoma using non-toxic cephalin as a fusogenic lipid. We compared our proposed method with PEG-mediated fusion, the well-known conventional method. Human lymphoblastoid cells were fused with an F3B6 heteromyeloma cell line using cephalin or PEG as the fusogenic compound. The viability of the cells and their fusion rate were determined microscopically and hybridoma (antigen-specific and non-specific) production yield was calculated following HAT selection and screening. The fusion rates of cells in cephalin and PEG-mediated methods were comparable (25.9+/-5.73% versus 27.3+/-6.07%) while the viability of the cells immediately and after overnight incubation was obviously greater in the cephalin method than in the PEG (p<0.001). Our proposed cephalin-mediated cell fusion method is about five times more efficient than PEG in production of hybridoma clones; thus it may dismiss PEG as the most generalized fusogen in the future.
Golestani
Reza
R
Department of Immunology, Tarbiat Modarres University, Tehran, Iran.
Pourfathollah
Ali Akbar
AA
Moazzeni
Seyed Mohammad
SM
D000818:Animals; D002417:Cattle; D002459:Cell Fusion; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D006825:Hybridomas / Q000187:drug effects; D051379:Mice; D010714:Phosphatidylethanolamines / Q000633:toxicity; D011092:Polyethylene Glycols / Q000633:toxicity; D012452:Saguinus
D010714:Phosphatidylethanolamines; D011092:Polyethylene Glycols