Cell culture and gene transcription effects of copper sulfate on Chinese hamster ovary cells
Cell culture and gene transcription effects of copper sulfate on Chinese hamster ovary cells
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硫酸铜对中国仓鼠卵巢细胞的细胞培养及基因转录影响
DOI：
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发表时间：
2011
期刊：
影响因子：
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通讯作者：
Z. Li
中科院分区：
文献类型：
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作者：
Yueming Qian;Sarwat Khattak;Zizhuo Xing;Aiqing He;P. Kayne;N. Qian;Shih;Z. Li
本文报道了不同浓度的硫酸铜对重组中国仓鼠卵巢(CHO)细胞系代谢和基因转录的影响。添加50μM硫酸铜可显著减少乳酸在培养物中的积累，同时提高活细胞密度和蛋白质效价。这些变化从第6天开始就可以看到，并随着培养时间的延长而变得越来越明显。将硫酸铜浓度降至5μM保留了上述所有有益效果，但增加了降低B0蛋白聚集态水平的好处。为了在转录水平上描述添加硫酸铜引起的细胞变化，Affymetrix®CHO微阵列被用来寻找与减少细胞压力和促进细胞周期相关的差异表达基因。基于芯片的结果，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多肽的上调被Western blotting证实。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关键作用，并强调了微阵列数据对于更好地了解对介质修改的生物反应的有用性。©2011美国化学工程师学会生物技术。节目，2011年
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 B0 protein. To profile the cellular changes due to copper sulfate addition at the transcriptional level, Affymetrix® 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. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011
Cu(II) 与铜蓝蛋白和牛血清白蛋白中非修复位点的结合。
影响因子：
3.9
作者：
Zgirski,A;Frieden,E
通讯作者：
Frieden,E
影响因子：
4.2
作者：
Prohaska, JR;Gybina, AA
通讯作者：
Gybina, AA