Affiliations: [a] Lehrstuhl für Biotechnologie, Universität Würzburg, D-8700 Würzburg, Germany | [b] Department of Veterinary Sciences, University of Arizona, Tucson, AZ, 85721, USA | [c] Department of Internal Medicine, University of Iowa, Iowa City, IA, 52242, USA
Note: [] Supported by the Deutsche Forschungsgemeinschaft (SFB 176, B5) and Federal Ministry for Research and Technology (DLR 01QV354) to UZ and NASA grant 7-816 to DWS and GAN.
Note: [] Address requests for reprints to Garry A. Neil, M.D., Department of Internal Medicine, 4612 JCP UIHC, University of Iowa, Iowa City, IA 52242, USA.
Abstract: We have developed and tested a novel electrofusion chamber, the adjustable plate microchamber, that permits the successful electrofusion and production of hybridomas in a hanging droplet from as few as 1,000 B lymphocytes. Cell suspension volumes of 10 μl may be used without excessive difficulty in aseptically recovering fused cells. With a modification of the hypo-osmolar electrofusion protocol with this microchamber, fusion efficiencies of the order of 10−3 may be attained. These efficiencies are comparable to those attained with standard hardware and much higher numbers of input B lymphocytes. This technology should permit high efficiency hybridoma production using a variety of hitherto inaccessible B lymphocyte populations such as B cells isolated from human organ biopsies.
