Cell hybridization by electrofusion on filters.
Electric field pulses induce permeabilization and associated fusogenicity in cell membranes. Electrofusion of cells is usually performed in two steps: the first is the creation of close intercellular contacts; the second is an application of electric pulses that induces membrane fusion. Very large cell contacts can be obtained by a filter aspiration method. A cell monolayer is created by controlled suction on biocompatible filter. No spontaneous fusion results. Just after filtration, electrofusion is obtained by field pulses applied parallel to the filter. Cell viability is not strongly affected and cells recover their spherical shape in the minute time range after filtration. The electrical parameters, the cell density, and the flow rate control fusion. Fusion is obtained with cells of different origins with very different adhesion properties. Hybrid cells are easily formed. This approach appears to be a very efficient method for cell hybridization with an easy-to-use protocol.
Ramos
Corinne
C
Institut de Pharmacologie et de Biologie Structurale, CNRS UMR 5089, 205, Route de Narbonne, 31077 Toulouse Cedex, France.
Bonenfant
Deborah
D
Teissie
Justin
J
D000818:Animals; D016466:CHO Cells / Q000166:cytology / Q000502:physiology; D002448:Cell Adhesion / Q000502:physiology; D002450:Cell Communication / Q000502:physiology; D002452:Cell Count; D002459:Cell Fusion / Q000379:methods*; D002463:Cell Membrane Permeability / Q000502:physiology; D048429:Cell Size / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D006224:Cricetinae; D004558:Electric Stimulation; D005374:Filtration / Q000379:methods; D006367:HeLa Cells / Q000166:cytology / Q000502:physiology; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology* / Q000502:physiology; D007939:Leukemia L1210 / Q000473:pathology; D008561:Membrane Fusion / Q000502:physiology*; D051379:Mice; D014407:Tumor Cells, Cultured