Antibodies, A Laboratory Manual, Second Edition, Edited by Edward A. Greenfield



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Ridding Cell Lines of Contaminating Microorganisms by Passage through Mice

(Protocol summary only for purposes of this preview site)

This method is for gross contamination of cultures and will work with the contents of a single microwell. Therefore, it can be used to rescue a relatively small culture. It will be necessary to single-cell-clone immediately after collecting the ascites. To use this technique, you will also need to have a screening assay available to identify the hybridoma cells from other cell types that will grow out. If the hybridoma cells form a solid ascitic tumor rather than a pocket of loose cells, it is still possible to rescue the cell line using this technique. The hybridoma cells will quickly grow out of tumor fragments dispersed in tissue culture medium. When mice are injected with contaminated hybridoma cells, they should be isolated from the remainder of the mouse colony to lessen the chances of infecting the other mice. Should the mice become ill from the infection, it will be necessary to treat the mice with antibiotics in their water supply. This method has a success rate of 80.

Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition

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