1H NMR Spectroscopy Profiling of Metabolic Reprogramming of Chinese Hamster Ovary Cells upon a Temperature Shift during Culture

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.