Electroacoustic production of murine hybridomas.
The optimal conditions for the production of murine hybridomas by electroacoustic fusion of cells in sugar solutions and of cells in ionic strength media (up to 115 mM NaCl) have been investigated. In the electroacoustic fusion technique cells were brought into contact in a 3 MHz ultrasonic standing wave field and were fused by application of an electric pulse. Hybridomas of murine splenocytes and X63-Ag8.653 mouse myelomas were successfully produced in low ionic strength mannitol solutions and in a range of salt concentrations up to 115 mM. The yield of hybridomas by electroacoustic fusion of cells in mannitol was at least as good as that obtained when cells were fused following conventional dielectrophoresis induced contact. The electroacoustic fusion yield was also comparable to conventional yields when cells were exposed to a pulse in higher ionic strength media where dielectrophoresis induces heating effects. Hybridomas were produced at similar electric field strengths when cells were suspended in high ionic strength media (up to 115 mM NaCl) or in mannitol. The effective electric field strength for hybridoma production was close to that at which trypan blue tests indicated membrane damage.
Bardsley
D W
DW
School of Pure and Applied Biology, University of Wales College of Cardiff, Cathays Park, U.K.
Liddell
J E
JE
Coakley
W T
WT
Clarke
D J
DJ
D000818:Animals; D002459:Cell Fusion; D004558:Electric Stimulation; D006825:Hybridomas / Q000276:immunology; D008722:Methods; D051379:Mice; D008807:Mice, Inbred BALB C; D009994:Osmolar Concentration; D013154:Spleen / Q000276:immunology; D014343:Trypan Blue; D014407:Tumor Cells, Cultured; D014465:Ultrasonics
D014343:Trypan Blue