Mammalian cell culture platform has been successfully implemented for industrial biopharmaceutical production through the advancements in early stage process development including cell-line engineering, media design and process optimization. However, late stage developments such as scale-up, scale-down and large-scale cell cultivation still face many industrial challenges to acquire comparable process performance between different culture scales. One of them is the sparging strategy which significantly affects productivity, quality and comparability. Currently, it is mainly relying on the empirical records due to the lack of theoretical framework and scarcity of available literatures to elucidate intracellular metabolic features. Therefore, it is highly required to characterize the underlying mechanism of physiological changes and metabolic states upon the aeration stress. To this end, initially we cultivated antibody producing CHO cells under mild and harsh sparging conditions and observed that sparging stress leads to the decreased cell growth rate, viability and productivity. Subsequent in silicomodel-driven flux analysis suggested that sparging stress rewires amino acid metabolism towards the enriched H2O2 turnover rate by up-regulated fluxes of amino acid oxidases. Interestingly, many of these H2O2-generating reactions were closely connected with the production of NADH, NADPH and GSH which are typical reducing equivalents. Thus, we can hypothesize that increased amino acid uptake caused by sparging stress contributes to restore redox homeostasis against oxidative stress. The current model-driven systematic data analysis allows us to quickly define distinct metabolic feature under stress condition by using basic cell cultivation datasets.

中文翻译：

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基于计算机模型的分批补料CHO细胞培养物中对剧烈刺激应激的代谢反应表征。

哺乳动物细胞培养平台已通过包括细胞系工程，培养基设计和工艺优化在内的早期工艺开发的成功实施，成功用于工业生物制药生产。然而，后期开发，例如按比例放大，按比例缩小和大规模细胞培养仍面临许多工业挑战，以在不同培养规模之间获得可比的工艺性能。其中之一是喷射策略，该策略会显着影响生产率，质量和可比性。目前，由于缺乏理论框架和缺乏可用于阐明细胞内代谢特征的现有文献，它主要依靠经验记录。因此，迫切需要表征曝气压力下生理变化和代谢状态的潜在机制。为此，最初，我们在温和苛刻的鼓泡条件下培养了产生抗体的CHO细胞，并观察到鼓泡应激会导致细胞生长速率，活力和生产率下降。随后的计算机模型驱动的通量分析表明，剧烈的应力通过氨基酸氧化酶通量的上调将氨基酸代谢重新导向富集的H2O2周转率。有趣的是，许多产生H2O2的反应与典型还原当量的NADH，NADPH和GSH的产生紧密相关。因此，我们可以假设由激增应激引起的氨基酸摄取增加有助于恢复氧化还原稳态以抵抗氧化应激。