¹H NMR spectroscopy profiling of metabolic reprogramming of Chinese hamster ovary cells upon a temperature shift during culture.
¹H NMR spectroscopy profiling of metabolic reprogramming of Chinese hamster ovary cells upon a temperature shift during culture.
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DOI：
10.1371/journal.pone.0077195
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Howard MJ
中科院分区：
文献类型：
--
作者：
Wagstaff JL;Masterton RJ;Povey JF;Smales CM;Howard MJ
我们报道了一种基于核磁共振的方法，通过研究CHOK1和CHO-S细胞在培养过程中以及对冷休克和随后的低温培养恢复的响应，来确定中国仓鼠卵巢细胞在培养过程中温度变化时的代谢重编程。CHOK1共鉴定出24种成分，CHO-S细胞系统鉴定出29种成分，其中CHO-S培养液中的葡萄糖含量是CHOK1培养液零时葡萄糖含量的5.6倍。我们证实，核磁共振代谢方法提供了葡萄糖和丙氨酸等成分的定量分析，两种细胞系的反应方式相似，并与先前报道的数据相当。然而，对乳酸的分析证实了CHOK1和CHO-S之间的差异，并且对温度的反应重新编程是细胞系特有的。我们的结果的意义是用主成分分析(PCA)来证实代谢物随温度和恢复的变化。最终，我们的方法证明了核磁共振提供实时分析的能力，以检测细胞对冷休克/亚生理温度的感知时代谢的重新编程。这有可能允许操纵培养上清液中的代谢物，以提高生长或生产率。
We report an NMR based approach to determine the metabolic reprogramming of Chinese hamster ovary cells upon a temperature shift during culture by investigating the extracellular cell culture media and intracellular metabolome of CHOK1 and CHO-S cells during culture and in response to cold-shock and subsequent recovery from hypothermic culturing. A total of 24 components were identified for CHOK1 and 29 components identified for CHO-S cell systems including the observation that CHO-S media contains 5.6 times the level of glucose of CHOK1 media at time zero. We confirm that an NMR metabolic approach provides quantitative analysis of components such as glucose and alanine with both cell lines responding in a similar manner and comparable to previously reported data. However, analysis of lactate confirms a differentiation between CHOK1 and CHO-S and that reprogramming of metabolism in response to temperature was cell line specific. The significance of our results is presented using principal component analysis (PCA) that confirms changes in metabolite profile in response to temperature and recovery. Ultimately, our approach demonstrates the capability of NMR providing real-time analysis to detect reprogramming of metabolism upon cellular perception of cold-shock/sub-physiological temperatures. This has the potential to allow manipulation of metabolites in culture supernatant to improve growth or productivity.
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