Genetic characterization of CHO production host DG44 and derivative recombinant cell lines

Derouazi, Madiha; Martinet, Danielle; Schmutz, Nathalie Besuchet; Flaction, R.; Wicht, M.; Bertschinger, Martin; Hacker, David L.; Beckmann, Jacques S.; Wurm, Florian Μ.

doi:10.1016/j.bbrc.2005.12.111

Cited by 135 publications

(110 citation statements)

References 21 publications

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“…Numerous studies have shown large chromosomal rearrangements in CHO cells, using banding techniques [41][42][43] and fluorescence in situ hybridization 8,[44][45][46][47][48] . These approaches identified large translocations in CHO cells, providing a coarse-grained view of genomic variations in these unstable genomes.…”

Section: Discussionmentioning

confidence: 99%

Genomic landscapes of Chinese hamster ovary cell lines as revealed by the Cricetulus griseus draft genome

Lewis¹,

et al. 2013

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Chinese hamster ovary (CHO) cells, first isolated in 1957, are the preferred production host for many therapeutic proteins. Although genetic heterogeneity among CHO cell lines has been well documented, a systematic, nucleotide-resolution characterization of their genotypic differences has been stymied by the lack of a unifying genomic resource for CHO cells. Here we report a 2.4-Gb draft genome sequence of a female Chinese hamster, Cricetulus griseus, harboring 24,044 genes. We also resequenced and analyzed the genomes of six CHO cell lines from the CHO-K1, DG44 and CHO-S lineages. This analysis identified hamster genes missing in different CHO cell lines, and detected >3.7 million single-nucleotide polymorphisms (SNPs), 551,240 indels and 7,063 copy number variations. Many mutations are located in genes with functions relevant to bioprocessing, such as apoptosis. The details of this genetic diversity highlight the value of the hamster genome as the reference upon which CHO cells can be studied and engineered for protein production.

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Section: Discussionmentioning

confidence: 99%

Genomic landscapes of Chinese hamster ovary cell lines as revealed by the Cricetulus griseus draft genome

Lewis¹,

et al. 2013

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“…As indicated above, one of the reasons for the varying results observed in the aforementioned studies is the high heterogeneity of CHO cells, which is in part due to their chromosomal instability and high rate of genomic rearrangements [11][12][13][14]. Two effects contribute: genomic variation between cell lines is observed [5], as well as what has been termed phenotypic drift: the slow but continuous changes and adaptations observed in cells maintained in culture for a prolonged period of time [15,16].…”

Section: Introductionmentioning

confidence: 99%

Microarray profiling of preselected CHO host cell subclones identifies gene expression patterns associated with in‐creased production capacity

Harreither

Hackl

Pichler

et al. 2015

Biotechnology Journal

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Over the last three decades, product yields from CHO cells have increased dramatically, yet specific productivity (qP) remains a limiting factor. In a previous study, using repeated cell-sorting, we have established different host cell subclones that show superior transient qP over their respective parental cell lines (CHO-K1, CHO-S). The transcriptome of the resulting six cell lines in different biological states (untransfected, mock transfected, plasmid transfected) was first explored by hierarchical clustering and indicated that gene activity associated with increased qP did not stem from a certain cellular state but seemed to be inherent for a high qP host line. We then performed a novel gene regression analysis identifying drivers for an increase in qP. Genes significantly implicated were first systematically tested for enrichment of GO terms using a Bayesian approach incorporating the hierarchical structure of the GO term tree. Results indicated that specific cellular components such as nucleus, ER, and Golgi are relevant for cellular productivity. This was complemented by targeted GSA that tested functionally homogeneous, manually curated subsets of KEGG pathways known to be involved in transcription, translation, and protein processing. Significantly implicated pathways included mRNA surveillance, proteasome, protein processing in the ER and SNARE interactions in vesicular transport.

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“…Plasmid DNA pMYKEF1-EGFP-puro was described previously (Derouazi et al 2006). Plasmid DNA was purified on a Nucleobond AX anion exchange column (Macherey-Nagel, Düren, Germany) according to the manufacturer's protocol.…”

Section: Cell Culturementioning

confidence: 99%

Suspension-adapted Chinese hamster ovary-derived cells expressing green fluorescent protein as a screening tool for biomaterials

et al. 2009

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Synthetic biomaterials play an important role in regenerative medicine. To be effective they must support cell attachment and proliferation in addition to being non-toxic and non-immunogenic. We used a suspension-adapted Chinese hamster ovary-derived cell line expressing green fluorescent protein (GFP) to assess cell attachment and growth on synthetic biomaterials by direct measurement of GFP-specific fluorescence. To simplify operations, all cell cultivation steps were performed in orbitallyshaken, disposable containers. Comparative studies between this GFP assay and previously established cell quantification assays demonstrated that this novel approach is suitable for rapid screening of a large number of samples. Furthermore the utility of our assay system was confirmed by evaluation of cell growth on three polyvinylidene fluoride polymer scaffolds that differed in pore diameter and drawing conditions. The data presented here prove the general utility of GFP-expressing cell lines and orbital shaking technology for the screening of biomaterials for tissue engineering applications.

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Genetic characterization of CHO production host DG44 and derivative recombinant cell lines

Cited by 135 publications

References 21 publications

Genomic landscapes of Chinese hamster ovary cell lines as revealed by the Cricetulus griseus draft genome

Genomic landscapes of Chinese hamster ovary cell lines as revealed by the Cricetulus griseus draft genome

Microarray profiling of preselected CHO host cell subclones identifies gene expression patterns associated with in‐creased production capacity

Suspension-adapted Chinese hamster ovary-derived cells expressing green fluorescent protein as a screening tool for biomaterials

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