Microscale production of hybridomas by hypo-osmolar electrofusion.
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 microliters 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.
Zimmermann
U
U
Lehrstuhl für Biotechnologie, Universität Würzburg, Germany.
Klöck
G
G
Gessner
P
P
Sammons
D W
DW
Neil
G A
GA
D000818:Animals; D001402:B-Lymphocytes; D002459:Cell Fusion; D002478:Cells, Cultured; D015194:Diffusion Chambers, Culture; D005260:Female; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C