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首页> 外文期刊>Biotechnology Progress >Cell Culture and Gene Transcription Effects of Copper Sulfate on Chinese Hamster Ovary Cells
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Cell Culture and Gene Transcription Effects of Copper Sulfate on Chinese Hamster Ovary Cells

机译：硫酸铜对中国仓鼠卵巢细胞的细胞培养和基因转录效应

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摘要

This study reports the effects of varying concentrations of copper sulfate on the metabolic and gene transcriptional profile of a recombinant Chinese hamster ovary (CHO) cell line producing an immunoglobulin G (IgG)-fusion protein (B0). Addition of 50 μM copper sulfate significantly decreased lactate accumulation in the cultures while increasing viable cell density and protein titer. These changes could be seen from day 6 and became increasingly evident with culture duration. Reducing the copper sulfate concentration to 5 μM retained all the above beneficial effects, but with the added benefit of reduced levels of the aggregated form of the BO protein. To profile the cellular changes due to copper sulfate addition at the transcriptional level, AffymetrixR CHO microarrays were used to identify differentially expressed genes related to reduced cellular stresses and facilitated cell cycling. Based on the microarray results, down-regulation of the transferrin receptor and lactate dehydrogenase, and up-regulation of a cytochrome P450 family-2 polypeptide were then confirmed by Western blotting. These results showed that copper played a critical role in cell metabolism and productivity on recombinant CHO cells and highlighted the usefulness of microarray data for better understanding biological responses on medium modification.
机译：这项研究报告了不同浓度的硫酸铜对产生免疫球蛋白G（IgG）融合蛋白（B0）的重组中国仓鼠卵巢（CHO）细胞系代谢和基因转录谱的影响。添加50μM硫酸铜可显着降低培养物中乳酸的积累，同时增加可行的细胞密度和蛋白滴度。从第6天开始就可以看到这些变化，随着文化持续时间的增加，这些变化变得越来越明显。将硫酸铜浓度降低至5μM保留了所有上述有益效果，但增加的好处是降低了BO蛋白聚集形式的水平。为了分析由于在转录水平上添加硫酸铜而引起的细胞变化，AffymetrixR CHO微阵列用于鉴定与减少的细胞应激和促进细胞周期相关的差异表达基因。基于微阵列结果，然后通过蛋白质印迹证实了转铁蛋白受体和乳酸脱氢酶的下调，以及细胞色素P450家族2多肽的上调。这些结果表明，铜在重组CHO细胞的细胞代谢和生产力中起着至关重要的作用，并强调了微阵列数据对于更好地了解培养基修饰的生物学反应的有用性。

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