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. 2014 Jan;66(1):133-47.
doi: 10.1007/s10616-013-9545-7. Epub 2013 Feb 22.

Mouse cell line authentication

Jamie L Almeida  1 , Carolyn R Hill, Kenneth D Cole
Affiliations

Mouse cell line authentication

Jamie L Almeida et al. Cytotechnology. 2014 Jan.

Abstract

The scientific community has responded to the misidentification of human cell lines with validated methods to authenticate these cells; however, few assays are available for nonhuman cell line identification. We have developed a multiplex polymerase chain reaction assay that targets nine tetranucleotide short tandem repeat (STR) markers in the mouse genome. Unique profiles were obtained from seventy-two mouse samples that were used to determine the allele distribution for each STR marker. Correlations between allele fragment length and repeat number were determined with DNA Sanger sequencing. Genotypes for L929 and NIH3T3 cell lines were shown to be stable with increasing passage numbers as there were no significant differences in fragment length with samples of low passage when compared to high passage samples. In order to detect cell line contaminants, primers for two human STR markers were incorporated into the multiplex assay to facilitate detection of human and African green monkey DNA. This multiplex assay is the first of its kind to provide a unique STR profile for each individual mouse sample and can be used to authenticate mouse cell lines.

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Figures

Fig. 1
Fig. 1
Genetic profile of the NIH3T3 cell line using the mouse multiplex assay (1 ng DNA)
Fig. 2
Fig. 2
Human contaminant detected in NIH3T3 STR profile (1:1 ratio of DNA from NIH3T3 and HeLa cell lines)
Fig. 3
Fig. 3
Genetic profile of the RAW 264.7 cell line using the mouse multiplex assay (1 ng DNA)
Fig. 4
Fig. 4
A mixture of NIH3T3 and RAW264.7 mouse cell lines detected using the mouse multiplex assay
See this image and copyright information in PMC

References

    1. Almeida J, Hill C, Cole K. Authentication of African green monkey cell lines using human short tandem repeat markers. BMC Biotechnol. 2011;11:102. doi: 10.1186/1472-6750-11-102. - DOI - PMC - PubMed
    1. Barallon R, Bauer SR, Butler J, Capes-Davis A, Dirks WG, Elmore E, Furtado M, Kline MC, Kohara A, Los GV, MacLeod RA, Masters JR, Nardone M, Nardone RM, Nims RW, Price PJ, Reid YA, Shewale J, Sykes G, Steuer AF, Storts DR, Thomson J, Taraporewala Z, Alston-Roberts C, Kerrigan L (2010) Recommendation of short tandem repeat profiling for authenticating human cell lines, stem cells, and tissues. In Vitro Cell Dev Biol Anim 46:727–732 - PMC - PubMed
    1. Barnes L, Bentley C, Dickson A. Advances in animal cell recombinant protein production: GS-NS0 expression system. Cytotechnology. 2000;32:109–123. doi: 10.1023/A:1008170710003. - DOI - PMC - PubMed
    1. Bonhomme F, Guenet JL. The laboratory mouse and its wild relatives. In: Lyon MF, Rastan S, Brown SDM, editors. Genetic variants and strains of the laboratory mouse. 3. Oxford, UK: Oxford University Press; 1996. pp. 1537–1576.
    1. Brownstein M, Carpten J, Smith J. Modulation of non-templated nucleotide addition by Taq DNA polymerase: primer modifications that facilitate genotyping. Biotechniques. 1996;20:1004–1010. - PubMed