Enhanced recombinant protein production in CHO cell continuous cultures under growth-inhibiting conditions is associated with an arrested cell cycle in G1/G0 phase.
Enhanced recombinant protein production in CHO cell continuous cultures under growth-inhibiting conditions is associated with an arrested cell cycle in G1/G0 phase.
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DOI：
10.1371/journal.pone.0277620
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发表时间：
2022
期刊：
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
--
作者：
低温和丁酸钠（NaBu）是生物制药生产过程中CHO细胞培养中最常用的两种提高生产率的策略。虽然这两种方法改变了细胞生长和生产力之间相互关系的平衡，但我们并不完全了解它们的作用机制，而不仅仅是抑制细胞生长。在这里，我们使用连续培养来评估低温和NaBu补充对CHO细胞性能和基因表达谱的差异影响。我们发现，在生长抑制条件下细胞生产率的增加与细胞在G1/G 0期的停滞有关。转录组分析显示，低温和NaBu将细胞周期阻滞在G1/G 0期的分子机制通过不同的细胞周期检查点和调节子的失调而彼此不同。当这两种策略组合时，在响应低温和NaBu时观察到的模式中的个体转录组变化被保留，导致在G1/G 0期阻滞细胞周期的累加效应。本文提出的研究结果提供了关于CHO细胞生物加工过程中细胞周期调控及其对增加重组蛋白产量的影响的新的分子见解。该数据为工程化生产力增强的CHO细胞系用于连续生产提供了背景。
Low temperature and sodium butyrate (NaBu) are two of the most used productivity-enhancing strategies in CHO cell cultures during biopharmaceutical manufacturing. While these two approaches alter the balance in the reciprocal relationship between cell growth and productivity, we do not fully understand their mechanisms of action beyond a gross cell growth inhibition. Here, we used continuous culture to evaluate the differential effect of low temperature and NaBu supplementation on CHO cell performance and gene expression profile. We found that an increase in cell-productivity under growth-inhibiting conditions was associated with the arrest of cells in the G1/G0 phase. A transcriptome analysis revealed that the molecular mechanisms by which low temperature and NaBu arrested cell cycle in G1/G0 differed from each other through the deregulation of different cell cycle checkpoints and regulators. The individual transcriptome changes in pattern observed in response to low temperature and NaBu were retained when these two strategies were combined, leading to an additive effect in arresting the cell cycle in G1/G0 phase. The findings presented here offer novel molecular insights about the cell cycle regulation during the CHO cell bioprocessing and its implications for increased recombinant protein production. This data provides a background for engineering productivity-enhanced CHO cell lines for continuous manufacturing.
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影响因子：
2.9
作者：
Altamirano, C;Paredes, C;Gòdia, F
通讯作者：
Gòdia, F
富含甘氨酸的RNA结合蛋白，介导了对哺乳动物细胞生长的冷诱导抑制。
影响因子：
7.8
作者：
Nishiyama, H;Itoh, K;Kaneko, Y;Kishishita, M;Yoshida, O;Fujita, J
通讯作者：
Fujita, J
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DOI：
10.4049/jimmunol.1700886
发表时间：
2018-02-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Grist JT;Jarvis LB;Georgieva Z;Thompson S;Kaur Sandhu H;Burling K;Clarke A;Jackson S;Wills M;Gallagher FA;Jones JL
通讯作者：
Jones JL
影响因子：
3.8
作者：
Bi, JX;Shuttleworth, J;Ai-Rubeai, M
通讯作者：
Ai-Rubeai, M
影响因子：
3.2
作者：
Chen, Fei;Kou, Tianci;Tan, Wen-Song
通讯作者：
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