Method for reducing spontaneous mutation background of wild type human-rat hybridoma AL cells

Abstract

The invention discloses a method for reducing a spontaneous mutation background of wild type human-rat hybridoma AL cells. The method comprises the following steps: collecting wild type cells; combining with a fluorescent antibody; preparing a sampling suspension solution; sorting the cells; and preserving. The method is simple to operate; the whole process from the collection of the cells and the combination with specific fluorescent proteins to sorting only needs 4-5 hours; continuously-cultured un-mutant (CD59<+>) cells and mutant (CD59<->) cells can be obtained at the same time so that the efficiency of detecting CD59 gene mutation can be improved greatly. With the adoption of the method, the spontaneous mutation rate of the AL cells can be reduced and about 30 mutant cells exist in 105 survival cells and are reduced by two times as much as the spontaneous mutation after Panning, so that the experiment sensitivity of a mutation detection system is improved effectively.

Description

technical field [0001] The invention relates to the technical field of mammalian cell mutation detection, in particular to a method for reducing wild-type human-mouse hybridoma A L Methods for spontaneous mutational backgrounds in cells. Background technique [0002] Gene mutation is not only one of the key factors of cancer, but also an important basis for evaluating the genotoxicity of environmental pollutants. A L Cells are Chinese hamster CHO cells gly - The immortalized cells obtained by hybridizing the A mutant strain with normal human fibroblasts contain a complete set of Chinese hamster CHO-K1 chromosomes and a human chromosome 11. Human chromosome 11 can encode a variety of cell surface proteins, including CD59 cell surface antigen (formerly known as S1 antigen) encoded by CD59 gene (formerly known as MIC1 gene). Since wild-type A L When the CD59 gene of the cells is normally expressed, the CD59 antigen is formed on the cell surface. In the presence of rabbit s...

AI Technical Summary

Problems solved by technology

It can be seen from the above that the operation process of this method is cumbersome, and a cycle usually takes 3 weeks, especially the process of treating the plate with IgM and washing the mutant cells requires a certain degree of proficiency of the experimental operator.

At the same time, the special antibody used in the experiment—goat anti-mouse IgM antibody is relatively expensive and used in large quantities.

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