Project description:CHO cells grown in serum-free medium have an absolute requirement for putrescine supplementation due to their deficiency in activity of the enzyme arginase. Consequently, principally in this system, polyamines play a central but poorly-understood role in cell proliferation Cell growth arrest, mainly in the S- and G2/M phases was observed. We used differential gene expression data to investigate the impact of polyamine deprivation on the transcriptome of CHO cells
Project description:CHO cells have primarily been studied in bulk, assuming that these bulk samples are identical because of genetic clonality across the sample. In this study, we performed single-cell RNA sequencing on two clonal CHO-K1 cell populations with different stability phenotypes over a 90 day culture period.
Project description:Transcriptional profiling of CHO-K1 cells comparing to in-house serum-free and suspension adapted CHO-K1 cells in the exponential phase. Goal was to determine the effects of serum on CHO-K1 cells.
Project description:In biopharmaceutical production, Chinese hamster ovary (CHO) cells derived from Cricetulus griseus remain the most commonly used host cell for recombinant protein production, especially antibodies. Over the last decade in-depth multi-omics characterization of these CHO cells provided data for extensive cell line engineering and corresponding increases in productivity. exosomes, extracellular vesicles containing proteins and nucleic acids, are barely researched at all in CHO cells. Exosomes have been proven to be an ubiquitous mediator of intercellular communication and are proposed as new biopharmaceutical format for drug delivery, indicator reflecting host cell condition and anti-apoptotic factor in spent media. Here we sequenced non-coding RNA of Exosomes (EXO) and whole cell lysate (WCL) isolated from CHO-K1 Cell Cultures at different growth phases (logarithmic/exponential phase (log/exp), stationary phase (stat), as well as death phase at 80 % viability (80 % ) and 60 % viability (60 %)) via Lexogen Small RNA-Seq Library Prep Kit for Illumina on the Illumina MiSeq platform in PE mode 2 x 36nt.
Project description:CHO-K1 (wildtype) and CHO-K6 (stablely overexpressing human HER2) cells were terated with 10 ug/ml transtuzumab or pertuzumab and their combination for 24 h and then the whole gene expression was analyzed by cDNA array.
Project description:The majority of recombinant protein therapeutics are produced with Chinese Hamster Ovary (CHO) cells. Productivity depends on the initial cell line engineering in terms of integration site or choice of an appropriate promotor for the recombinant gene expression, as well as media and process parameter optimization. Here, proteomic profiling is used to identify optimization targets for a pharmaceutical relevant cell line system. Triplicates of CHO-K1 cell 2 L bioreactor fedbatch cultivations were performed and daily sampled for nLC-MS/MS proteomic analysis. Collected data from day 3 up to day 11 showed high Pearson correlation of 93.7 ± 4 % with ca. 2500 proteins quantified. The different growth phases were separated by principal component analysis and hierarchical clustering approaches. Subsequent statistical analysis revealed distinct protein profiles, where steady increase or decrease over time were the most prominent clusters. Fisher exact enrichment tests yielded in significantly enriched protein annotations which were successfully mapped to growth and metabolic changes during fedbatch cell cultivation. Major improvements in cellular and process understanding were achieved and yielded in the identification of promising new targets, like strong endogenous promotors, for cellular engineering and process optimization of this biopharmaceutical relevant cell line.