1. Genetic characterization of CHO production host DG44 and derivative recombinant cell lines
- Author
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Florian M. Wurm, R. Flaction, Danielle Martinet, Martin Bertschinger, Madiha Derouazi, Jacques S. Beckmann, David L. Hacker, N. Besuchet Schmutz, M. Wicht, Laboratory of Cellular Biotechnology [Lausanne] (LBTC), IGBB - Institute of Biological Engineering and Biotechnology, Groupe de Recherche et d'Etude du Processus Inflammatoire (GREPI), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Laboratory of Cytogenetics, Centre Hospitalier Universitaire Vaudois [Lausanne] (CHUV), and This work was supported by Pfizer Inc.
- Subjects
0106 biological sciences ,[SDV.BIO]Life Sciences [q-bio]/Biotechnology ,MESH: Chromosomes, Mammalian ,DNA Mutational Analysis ,Variation (Genetics) ,Aneuploidy ,MESH: Cricetinae ,01 natural sciences ,Biochemistry ,CHO Cells/*metabolism ,law.invention ,MESH: Recombinant Proteins ,chemistry.chemical_compound ,MESH: Cricetulus ,law ,Cricetinae ,MESH: Animals ,MESH: Genetic Variation ,MESH: DNA Mutational Analysis ,[INFO.INFO-BT]Computer Science [cs]/Biotechnology ,Calcium phosphate transfection ,Recombination, Genetic ,0303 health sciences ,Recombinant ,medicine.diagnostic_test ,Chinese hamster ovary cell ,Chromosome Mapping ,Karyotype ,Microinjection ,Recombinant Proteins ,Recombinant DNA ,MESH: Recombination, Genetic ,Chromosome Aberrations/*statistics & numerical data ,Ploidy ,Karyotype CHO DG44 ,Green Fluorescent Proteins ,Biophysics ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,CHO Cells ,Recombinant Proteins/*biosynthesis/*genetics ,Biology ,GFP ,Chromosomes ,Chinese hamster ,03 medical and health sciences ,MESH: Green Fluorescent Proteins ,Cricetulus ,FISH ,MESH: CHO Cells ,010608 biotechnology ,Green Fluorescent Proteins/*genetics ,medicine ,MESH: Chromosome Aberrations ,Animals ,Molecular Biology ,Genetic/genetics ,030304 developmental biology ,Chromosome Aberrations ,[SDV.GEN]Life Sciences [q-bio]/Genetics ,Genetic Variation ,Cell Biology ,Mammalian/*genetics/ultrastructure ,medicine.disease ,biology.organism_classification ,Molecular biology ,Chromosomes, Mammalian ,Recombination ,chemistry ,MESH: Chromosome Mapping ,DNA ,Fluorescence in situ hybridization - Abstract
International audience; The dihydrofolate reductase-deficient Chinese hamster ovary (CHO) cell line DG44 is the dominant mammalian host for recombinant protein manufacturing, in large part because of the availability of a well-characterized genetic selection and amplification system. However, this cell line has not been studied at the cytogenetic level. Here, the first detailed karyotype analysis of DG44 and several recombinant derivative cell lines is described. In contrast to the 22 chromosomes in diploid Chinese hamster cells, DG44 has 20 chromosomes, only seven of which are normal. In addition, four Z group chromosomes, seven derivative chromosomes, and 2 marker chromosomes were identified. For all but one of the 16 DG44-derived recombinant cell lines analyzed, a single integration site was detected by fluorescence in situ hybridization regardless of the gene delivery method (calcium phosphate-DNA coprecipitation or microinjection), the topology of the DNA (circular or linear), or the integrated plasmid copy number (between 1 and 51). Chromosomal aberrations, observed in more than half of the cell lines studied, were mostly unbalanced with examples of aneuploidy, deletions, and complex rearrangements. The results demonstrate that chromosomal aberrations are frequently associated with the establishment of recombinant CHO DG44 cell lines. Noteworthy, there was no direct correlation between the stability of the genome and the stability of recombinant protein expression.
- Published
- 2005
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